3-0x0-2,3-dihydro-1h-isoindole-4-carboxamides with selective parp-1 inhibition

ABSTRACT

There are provided substituted 3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide derivatives (I) which selectively inhibit the activity of poly (ADP-ribose) polymerase PARP-1 with respect to poly (ADP-ribose) polymerase PARP-2. The compounds of this invention are therefore useful in treating diseases such as cancer, cardiovascular diseases, central nervous system injury and different forms of inflammation. The present invention also provides methods for preparing these compounds, pharmaceutical compositions comprising these compounds, and methods of treating diseases utilizing pharmaceutical compositions comprising these compounds.

The present invention provides substituted3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide derivatives whichselectively inhibit the activity of poly (ADP-ribose) polymerase PARP-1with respect to poly (ADP-ribose) polymerase PARP-2. The compounds ofthis invention are therefore useful in treating diseases such as cancer,cardiovascular diseases, central nervous system injury and differentforms of inflammation. The present invention also provides methods forpreparing these compounds, pharmaceutical compositions comprising thesecompounds, and methods of treating diseases utilizing pharmaceuticalcompositions comprising these compounds.

Poly (ADP-ribose) polymerases belong to a family of 18 members thatcatalyze the addition of ADP-ribose units to DNA or different acceptorproteins, which affect cellular processes as diverse as replication,transcription, differentiation, gene regulation, protein degradation andspindle maintenance. PARP-1 and PARP-2 are the only enzymes among thePARPs that are activated by DNA damage and are involved in DNA repair.

PARP-1 is a nuclear protein consisting of three domains: the N-terminalDNA-binding domain containing two zinc fingers, the auto modificationdomain, and the C-terminal catalytic domain. PARP-1 binds through thezinc-finger domain to DNA single strand breaks (SSB), cleaves NAD+, andattaches multiple ADP-ribose units to target proteins such as histonesand various DNA repair enzymes. This results in a highly negativelycharged target, which in turn leads to the unwinding and repair of thedamaged DNA through the base excision repair pathway. In knock out mousemodels, deletion of PARP-1 impairs DNA repair but it is not embryoniclethal. Double knock out PARP-1 and PARP-2 mice instead die during earlyembryogenesis, suggesting that the two enzymes display not completelyoverlapping functions. Enhanced PARP-1 expression and/or activity havebeen shown in different tumor cell lines, including malignant lymphomas,hepatocellular carcinoma, cervical carcinoma, colorectal carcinoma,leukemia. This may allow tumor cells to withstand genotoxic stress andincrease their resistance to DNA-damaging agents. As a consequence,inhibition of PARP-1 through small molecules has been shown to sensitizetumor cells to cytotoxic therapy (e.g. temozolomide, platinums,topoisomerase inhibitors and radiation). A significant window seems toexist between the ability of a PARP inhibitor to potentiate therapeuticbenefits and undesirable side effects. Whereas the therapeutic use ofPARP inhibitors in combination with DNA damaging agents is not novel,the use of these agents as monotherapy, in particular tumor geneticbackgrounds deficient in the homologous recombination DNA repair,represents a new approach. Individuals with heterozygous germ linemutations in either the BRCA-1 or BRCA-2 homologous recombination repairgenes exhibit high life time risks of developing breast and othercancers. Tumors arising in mutation carriers have generally lost thewild type allele and do not express functional BRCA-1 and BRCA-2proteins.

Therefore, loss of these two proteins leads to a tumor-specificdysfunction in the repair of double strand breaks by homologousrecombination. It is known that when PARP-1 is inhibited, base excisionrepair is reduced and single strand breaks that are generated during thenormal cell cycle persist. It has also been established that replicationforks that encounter an unrepaired break can form double strand breakswhich are normally repaired by homologous recombination. Tumor cellsthat are deficient in homologous recombination repair such as BRCA-1 andBRCA-2 mutants are therefore highly sensitive to PARP inhibitioncompared with wild-type cells. This is in line with the concept ofsynthetic lethality, in which the two pathway defects alone areinnocuous but combined become lethal: PARP inhibitors may be moreeffective in patients with tumors with specific DNA repair defectswithout affecting normal heterozygous tissues. Putative patientpopulation includes, besides BRCA mutants that represent the majority ofhereditary breast and ovarian cancer, also a substantial fraction ofsporadic cancers with defects in homologous recombination repair, aphenomenon termed “BRCAness”. For example, methylation of the promotersof the BRCA-1 or FANCF genes and amplification of the EMSY gene, whichencodes a BRCA-2 interacting protein. By extending the rational ofsynthetic lethality of PARP and BRCA-1 and BRCA-2, it is likely thatdeficiencies in any gene that is not redundant in double strand breakrepair should be sensitive to PARP inhibition. For example, ATMdeficiency, found in patients with T-cell prolymhocytic leukemia andB-cell chronic lymphocitic leukemia and breast cancer and CHK2 germ linemutations identified in sarcoma, breast cancer, ovarian cancer and braintumors, have also been shown to be synthetically lethal in combinationwith PARP deficiency as well as deficiencies in other known HR pathwayproteins (including RAD51, DSS1, RAD54, RPA1, NBS1, ATR, CHK1, CHK2,FANCD2, FANCA, and FANCC).

Mutations in FANCC and FANCG have been shown in pancreatic cancer.Methylation of FANCF promoter has been found in ovarian, breast,cervical, lung carcinomas. The first clinical evidence that BRCA-mutatedcancer may be sensitive to PARP inhibitor monotherapy comes from thepreliminary data for the phase I trial of the oral, small molecule PARPinhibitor AZD2281. In an enriched phase I population for BRCA mutationcarriers, partial responses were seen in 4 out of 10 ovarian cancerpatients with confirmed BRCA-1 mutations. Other PARP inhibitors, such asAG014699, BSI-201, are currently known to be in phase II clinical trialsboth in combination with DNA damaging agents and as single agent in BRCAdeficient tumors. Early indications are that these therapies show lowtoxicity. Anyway compounds with high selectivity on PARP-1 are expectedto show even less toxicity in view of a chronic treatment schedule.

PARP-1 has also been implicated in angiogenesis. In particular, PARP-1inhibition seems to result in decreased accumulation of thetranscription hypoxia-inducible factor 1α, an important regulator oftumor cell adaptation to hypoxia.

Pro-inflammatory stimuli trigger the release of pro-inflammatorymediators that induce the production of peroxynitrate and hydroxylradicals, which in turn yield to DNA single strand breakage withconsequent activation of PARP-1. Over activation of PARP-1 results indepletion of NAD+ and energy stores, culminating in cell dysfunction andnecrosis. This cellular suicide mechanism has been implicated in thepathomechanism of stroke, myocardial ischemia, diabetes,diabetes-associated cardiovascular dysfunction, shock, traumatic centralnervous system injury, arthritis, colitis, allergic encephalomyelitisand various other forms of inflammation. Of special interest is theenhancement by PARP-1 of nuclear factor kB-mediated transcription, whichplays a central role in the expression of inflammatory cytokines,chemokines and inflammatory mediators.

WO2007047646 in the name of Janssen Pharmaceutica describes substituteddihydro isoindolones useful for treating kinase disorders; Wender et al.claim in U.S. Pat. No. 7,232,842 isoindolone analogs as kinaseinhibitors. The Patent Application Publication US 2008/0108659 of Gandhiet al. describes 3-oxo-2,3-dihydro-1H-isoindoles as poly (ADP-Ribose)polymerase inhibitors, also reported in: Bioorg. Med. Chem. Lett., 2010,20, 1023-1026. The present invention provides novel 1H-isoindolederivatives which are selective PARP-1 inhibitors with respect to PARP-2and are thus useful in the therapy of cancer, cardiovascular diseases,nervous system injury and inflammation.

Accordingly, a first object of the present invention is to provide acompound of the formula (I):

whereinR is an optionally substituted linear or branched C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl orheteroaryl group;R₁ and R₂ are independently hydrogen atom, an optionally substitutedlinear or branched C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇cycloalkyl, heterocyclyl, aryl or heteroaryl group, or, taken togetherwith the ring carbon atom to which they are bonded, form an optionallysubstituted carbon-carbon double bond, a C₃-C₇ cycloalkyl orheterocyclyl group, with the proviso that R₁ and R₂ are not bothhydrogen atoms;R₃ is hydrogen or halogen atom, cyano, nitro or an optionallysubstituted linear or branched C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl, NHCOR₄, COR₄,NR₅R₆, NR₅COR₄, OR₇, SR₇, SOR₁₀, SO₂R₁₀, NHSOR₁₀, NHSO₂R₁₀, R₈R₉N—C₁-C₆alkyl or R₈O—C₁-C₆ alkyl group;R₄ is hydrogen atom or an optionally substituted linear or branchedC₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl, NR₅R₇,OR₇, SR₇, R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl group;R₅ and R₆ are independently hydrogen atom, or an optionally substitutedlinear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, heterocyclyl, aryl, heteroaryl, R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆alkyl group, or R₅ and R₆, taken together with the nitrogen atom towhich they are bonded, form an optionally substituted heterocyclylgroup;R₇ is hydrogen atom, or an optionally substituted linear or branchedC₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,heterocyclyl, aryl, heteroaryl, COR₅, SOR₁₀, SO₂R₁₀, R₈R₉N—C₁-C₆ alkyl,R₈O—C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkyl orheteroaryl-C₁-C₆ alkyl group, wherein R₅ is as defined above;R₈ and R₉ are independently hydrogen atom or an optionally substitutedlinear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, heterocyclyl, aryl, heteroaryl, COR₄, or R₈ and R₉, takentogether with the nitrogen atom to which they are bonded, form anoptionally substituted heterocyclyl group, wherein R₄ is as definedabove;R₁₀ is hydrogen atom, or an optionally substituted linear or branchedC₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,heterocyclyl, aryl, heteroaryl, NR₅R₆, OR₇, R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆alkyl, wherein R₅, R₆, R₇, R₈ and R₉ are as defined above;or a pharmaceutically acceptable salt thereof.

The compounds of formula (I) are potent and selective PARP-1 inhibitorswith respect to PARP-2 and are thus useful in cancer, cardiovasculardiseases, nervous system injury and inflammation therapy.

The present invention also provides methods of synthesizing3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide derivatives of formula (I)prepared through a process consisting of standard synthetictransformations.

The present invention also provides methods for treating diseasesmediated by the PARP-1 protein.

A preferred method of the present invention is to treat a diseasemediated by PARP-1 protein selected from the group consisting of cancer,cardiovascular diseases, nervous system injury and inflammation.

Another preferred method of the present invention is to treat specifictypes of cancer including but not limited to: carcinoma such as bladder,breast, colon, kidney, liver, lung, including small cell lung cancer,esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid,prostate, and skin, including squamous cell carcinoma; hematopoietictumors of lymphoid lineage including leukaemia, acute lymphocyticleukaemia, acute lymphoblastic leukaemia, B-cell lymphoma,T-cell-lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy celllymphoma and Burkitt's lymphoma; hematopoietic tumors of myeloidlineage, including acute and chronic myelogenous leukemias,myelodysplastic syndrome and promyelocytic leukaemia; tumors ofmesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; tumorsof the central and peripheral nervous system, including astrocytoma,neuroblastoma, glioma and schwannomas; other tumors, including melanoma,seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum,keratoxanthoma, thyroid follicular cancer and Kaposi's sarcoma.

Another preferred method of the present invention is to treat specifictypes of cardiovascular diseases including but not limited to:myocardial reperfusion injury, cardiomyopathy, diabetic cardiovasculardysfunction.

Another preferred method of the present invention is to treat specifictypes of central nervous system injury including but not limited tostroke, brain injury and neurodegenerative disorders.

The present invention further provides a method of treatment comprisinga compound of formula (I) in combination with radiation therapy orchemotherapy regimen for simultaneous, separate or sequential use inanticancer therapy. Moreover the invention provides an in vitro methodfor selectively inhibiting the PARP-1 protein activity which comprisescontacting the said protein with an effective amount of a compound offormula (I).

The present invention also provides a pharmaceutical compositioncomprising one or more compounds of formula (I) or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable excipient,carrier or diluent.

The present invention also provides a pharmaceutical compositioncomprising a compound of formula (I) in combination with knowncytostatic or cytotoxic agents, antibiotic-type agents, alkylatingagents, antimetabolite agents, hormonal agents, immunological agents,interferon-type agents, cyclooxygenase inhibitors (e.g. COX-2inhibitors), matrix metalloprotease inhibitors, telomerase inhibitors,tyrosine kinase inhibitors, anti-growth factor receptor agents, anti-HERagents, anti-EGFR agents, anti-angiogenesis agents (e.g. angiogenesisinhibitors), farnesyl transferase inhibitors, ras-raf signaltransduction pathway inhibitors, cell cycle inhibitors, other cdksinhibitors, tubulin binding agents, topoisomerase I inhibitors,topoisomerase II inhibitors, and the like. Additionally, the inventionprovides a product or kit comprising a compound of formula (I) or apharmaceutically acceptable salt thereof, as defined above, orpharmaceutical compositions thereof and one or more chemotherapeuticagents, as a combined preparation for simultaneous, separate orsequential use in anticancer therapy.

In yet another aspect, the invention provides a compound of formula (I)or a pharmaceutically acceptable salt thereof, as defined above, for useas a medicament, preferably as a medicament with antitumor activity.

Moreover the invention provides the use of a compound of formula (I) ora pharmaceutically acceptable salt thereof, as defined above, in themanufacture of a medicament with antitumor activity.

Finally, the invention provides a compound of formula (I) or apharmaceutically acceptable salt thereof, as defined above, for use in amethod of treating cancer.

The compounds of formula (I) may have one or more asymmetric centers,and may therefore exist as individual optical isomers or racemicmixtures or diastereoisomer. Accordingly, all the possible isomers, andtheir mixtures of the compounds of formula (I) are within the scope ofthe present invention. As stated above, salts of the compounds offormula (I) are also within the scope of the present invention.

Unless otherwise specified, when referring to the compounds of formula(I) per se as well as to any pharmaceutical composition thereof or toany therapeutic treatment comprising them, the present inventionincludes all of the isomers, tautomers, hydrates, solvates, N-oxides andpharmaceutically acceptable salts of the compounds of this invention.

If a chiral center or another form of an isomeric center is present in acompound of the present invention, all forms of such isomer or isomers,including enantiomers and diastereomers, are intended to be coveredherein. Compounds containing a chiral center may be used as a racemicmixture, an enantiomerically enriched mixture, or the racemic mixturemay be separated using well-known techniques and an individualenantiomer may be used alone. In cases in which compounds haveunsaturated carbon-carbon double bonds, both the cis (Z) and trans (E)isomers are within the scope of this invention.

In cases wherein compounds may exist in tautomeric forms, such asketo-enol tautomers, each tautomeric form is contemplated as beingincluded within this invention whether existing in equilibrium orpredominantly in one form.

With the term halogen atom we intend a fluorine, chlorine, bromine oriodine atom.

With the term cyano we intend a —CN residue.

With the term nitro we intend a —NO₂ group.

With the term “linear or branched C₁-C₆ alkyl”, we intend any of thegroups such as, for instance, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, n-hexyl, and thelike.

With the term “C₂-C₆ alkenyl” we intend an aliphatic C₂-C₆ hydrocarbonchain containing at least one carbon-carbon double bond and which can belinear or branched. Representative examples include, but are not limitedto, ethenyl, 1-propenyl, 2-propenyl, 1- or 2-butenyl, and the like.

With the term “C₂-C₆ alkynyl” we intend an aliphatic C₂-C₆ hydrocarbonchain containing at least one carbon-carbon triple bond and which can belinear or branched. Representative examples include, but are not limitedto, ethynyl, 1-propynyl, 2-propynyl, 1- or 2-butynyl, and the like.

With the term “C₃-C₇ cycloalkyl” we intend, unless otherwise provided, a3- to 7-membered all-carbon monocyclic ring, which may contain one ormore double bonds but does not have a completely conjugated π-electronsystem. Examples of cycloalkyl groups, without limitation, arecyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane,cyclohexene and cyclohexadiene.

With the term “heterocyclyl” we intend a 3- to 8-membered, saturated orpartially unsaturated carbocyclic ring where one or more carbon atomsare replaced by heteroatoms such as nitrogen, oxygen and sulfur. Nonlimiting examples of heterocyclyl groups are, for instance, pyrane,pyrrolidine, pyrroline, imidazoline, imidazolidine, pyrazolidine,pyrazoline, thiazoline, thiazolidine, dihydrofuran, tetrahydrofuran,1,3-dioxolane, piperidine, piperazine, morpholine and the like.

The term “aryl” refers to a mono-, bi- or poly-carbocyclic hydrocarbonwith from 1 to 4 ring systems, optionally further fused or linked toeach other by single bonds, wherein at least one of the carbocyclicrings is “aromatic”, wherein the term “aromatic” refers to completelyconjugated π-electron bond system. Non limiting examples of such arylgroups are phenyl, α- or β-naphthyl or biphenyl groups.

The term “heteroaryl” refers to aromatic heterocyclic rings, typically5- to 8-membered heterocycles with from 1 to 3 heteroatoms selectedamong N, O or S; the heteroaryl ring can be optionally further fused orlinked to aromatic and non-aromatic carbocyclic and heterocyclic rings.Not limiting examples of such heteroaryl groups are, for instance,pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, imidazolyl,thiazolyl, isothiazolyl, pyrrolyl, phenyl-pyrrolyl, furyl, phenyl-furyl,oxazolyl, isoxazolyl, pyrazolyl, thienyl, benzothienyl, isoindolinyl,benzoimidazolyl, indazolyl, quinolinyl, isoquinolinyl, 1,2,3-triazolyl,1-phenyl-1,2,3-triazolyl, 2,3-dihydroindolyl, 2,3-dihydrobenzofuranyl,2,3-dihydrobenzothiophenyl; benzopyranyl, 2,3-dihydrobenzoxazinyl,2,3-dihydroquinoxalinyl and the like.

According to the present invention and unless otherwise provided, any ofthe above R-R₃ group may be optionally substituted, in any of their freepositions, by one or more groups, for instance 1 to 6 groups,independently selected from: halogen, nitro, oxo groups (═O), cyano,C₁-C₆ alkyl, polyfluorinated alkyl, polyfluorinated alkoxy, C₂-C₆alkenyl, C₂-C₆ alkynyl, hydroxyalkyl, aryl, arylalkyl, heteroaryl,heteroarylalkyl, heterocyclyl, heterocyclylalkyl, C₃-C₇ cycloalkyl,hydroxy, alkoxy, aryloxy, heterocyclyloxy, methylenedioxy,alkylcarbonyloxy, arylcarbonyloxy, cycloalkenyloxy,heterocyclylcarbonyloxy, alkylideneaminooxy, carboxy, alkoxycarbonyl,aryloxycarbonyl, cycloalkyloxycarbonyl,heterocyclylalkyloxycarbonyl-amino, ureido, alkylamino, dialkylamino,arylamino, diarylamino, heterocyclylamino, formylamino,alkylcarbonylamino, arylcarbonylamino, heterocyclylcarbonylamino,aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,arylaminocarbonyl, heterocyclylaminocarbonyl, alkoxycarbonylamino,hydroxyaminocarbonyl alkoxyimino, alkylsulfonylamino, arylsulfonylamino,heterocyclylsulfonylamino, formyl, alkylcarbonyl, arylcarbonyl,cycloalkylcarbonyl, heterocyclylcarbonyl, alkylsulfonyl, arylsulfonyl,aminosulfonyl, alkylaminosulfonyl, dialkylaminosulfonyl,arylaminosulfonyl, heterocyclylaminosulfonyl, arylthio, alkylthio,phosphonate and alkylphosphonate. In their turn, whenever appropriate,each of the above substituent may be further substituted by one or moreof the aforementioned groups.

With the term polyfluorinated alkyl or polyfluorinated alkoxy we intendany of the above linear or branched C₁-C₆ alkyl or alkoxy groups whichare substituted by more than one fluorine atom such as, for instance,trifluoromethyl, trifluoroethyl, 1,1,1,3,3,3-hexafluoropropyl,trifluoromethoxy and the like.

With the term hydroxyalkyl we intend any of the above C₁-C₆ alkyl,bearing an hydroxyl group such as, for instance, hydroxymethyl,2-hydroxyethyl, 3-hydroxypropyl and the like.

From all of the above, it is clear to the skilled person that any groupwhich name is a composite name such as, for instance, arylamino has tobe intended as conventionally construed by the parts from which itderives, e.g. by an amino group which is further substituted by aryl,wherein aryl is as above defined.

Likewise, any of the terms such as, for instance, alkylthio, alkylamino,dialkylamino, alkoxycarbonyl, alkoxycarbonylamino, heterocyclylcarbonyl,heterocyclylcarbonylamino, cycloalkyloxycarbonyl and the like, includegroups wherein the alkyl, alkoxy, aryl, C₃-C₇ cycloalkyl andheterocyclyl moieties are as above defined.

In particular, in the above definition, “optionally substituted” meansthat the group may be substituted with one or more substituentsindependently selected from linear or branched C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃-C₇ cycloalkyl-C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl,aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆ alkyl group, halogen, cyano, nitro,NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), SR₇, SOR₁₀, SO₂R₁₀,NHSOR₁₀, NHSO₂R₁₀, R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆ alkyl group.

All those substituents may be optionally further substituted with one ormore substituents independently selected from halogen, C₁-C₆ alkyl, arylor heterocyclyl group, NHCOR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), SR₇,R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆ alkyl, wherein R₄, R₅, R₆, R₇, R₈, R₉ andR₁₀ are as defined above.

The term “pharmaceutically acceptable salt” of compounds of formula (I)refers to those salts that retain the biological effectiveness andproperties of the parent compound, therefore pharmaceutically acceptablesalts of the compounds of formula (I) include the acid addition saltswith inorganic or organic acids, e.g., nitric, hydrochloric,hydrobromic, sulfuric, perchloric, phosphoric, acetic, trifluoroacetic,propionic, glycolic, (D) or (L) lactic, oxalic, ascorbic, fumaric,malonic, malic, maleic, tartaric, citric, benzoic, cinnamic, mandelic,methanesulphonic, ethanesulfonic, p-toluenesulfonic, isethionic,succinic and salicylic acid.

Pharmaceutically acceptable salts of the compounds of formula (I) alsoinclude the salts with inorganic or organic bases, e.g., alkali oralkaline-earth metals, especially sodium, potassium, calcium, ammoniumor magnesium hydroxides, carbonates or bicarbonates, acyclic or cyclicamines, preferably methylamine, ethylamine, diethylamine, triethylamine,piperidine and the like.

In a first preferred embodiment, compounds of general formula (I) arecharacterized in that R is an optionally substituted C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, the optional substituents beingone or more halogen, cyano, an optionally further substituted linear orbranched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl,C₃-C₇ cycloalkyl-C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkylor heteroaryl-C₁-C₆ alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo(═O), R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, the optional furthersubstituents being one or more halogen, C₁-C₆ alkyl, aryl orheterocyclyl group, NHCOR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), SR₇,R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, and

R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ have the meanings above defined,or a pharmaceutically acceptable salt thereof.

In a second preferred embodiment, compounds of general formula (I) arecharacterized in that R is an optionally substituted C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, the optional substituents beingone or more halogen, cyano, an optionally further substituted linear orbranched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroarylor C₃-C₇ cycloalkyl-C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆alkyl or heteroaryl-C₁-C₆ alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄,OR₇, oxo (═O), R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, the optionalfurther substituents being one or more halogen, C₁-C₆ alkyl, aryl orheterocyclyl group, NHCOR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), SR₇,R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl;

either R₁ is hydrogen atom, and R₂ is an optionally substituted linearor branched C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, orR₁ and R₂, taken together with the ring carbon atom to which they arelinked, form an optionally substituted carbon-carbon double bond, aC₃-C₇ cycloalkyl or heterocyclyl group, andR₃, R₄, R₅, R₆, R₇, R₈ and R₉ have the meanings above defined,or a pharmaceutically acceptable salt thereof.

In a third preferred embodiment, compounds of general formula (I) arecharacterized in that R is an optionally substituted C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, the optional substituents beingone or more halogen, cyano, an optionally further substituted linear orbranched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl,C₃-C₇ cycloalkyl-C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkylor heteroaryl-C₁-C₆ alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo(═O), R₈R₈N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, the optional furthersubstituents being one or more halogen, C₁-C₆ alkyl, aryl orheterocyclyl group, NHCOR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), SR₇,R₈R₈N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl group;

either R₁ is hydrogen atom, and R₂ is an optionally substituted linearor branched C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl or heteroaryl group,or R₁ and R₂, taken together with the ring carbon atom to which they arelinked, form an optionally substituted carbon-carbon double bond, aC₃-C₇ cycloalkyl or heterocyclyl group;R₃ is hydrogen, halogen atom, cyano, nitro, an optionally substitutedlinear or branched C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, NHCOR₄,NR₅R₆, NR₅COR₄, OR₇, R₈R₈N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, andR₄, R₅, R₆, R₇, R₈, and R₉ are as defined above,or a pharmaceutically acceptable salt thereof.

Even more preferably, the present invention provides compounds of theformula (I) as defined above characterized in that:

R is an optionally substituted heterocyclyl or aryl group, the optionalsubstituents being, one or more, optionally further substituted linearor branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃-C₇ cycloalkyl-C₁-C₆alkyl, heterocyclyl-C₁-C₆alkyl,aryl-C₁-C₆alkyl or heteroaryl-C₁-C₆alkyl group or COR₄, and, theoptional further substituents being one or more halogen, C₁-C₆ alkyl,aryl or heterocyclyl group, OR₇, oxo (═O), SR₇, or R₈O—C₁-C₆ alkylgroup;R₁ and R₂ are independently hydrogen atom or a C₁-C₆ alkyl group, withthe proviso that they are not both hydrogen atoms;R₃ is hydrogen or halogen atom;

R₄ is OR₇,

R₇ is hydrogen atom, optionally substituted C₁-C₆ alkyl or aryl-C₁-C₆alkyl group, substituents being one or more halogen atom;R₈ is hydrogen atom, andR₅, R₆ and R₉ are as defined above,or a pharmaceutically acceptable salt thereof.

Other preferred compounds provided from the present invention are thecompounds of the formula (I) as defined above characterized in that

R is an optionally substituted piperidinyl or phenyl group, the optionalsubstituents being, one or more, optionally further substituted methyl,ethyl, propyl, cyclohexyl, cyclopentyl, cyclobutyl, morpholinyl,piperazinyl, pyrazolyl, cyclohexyl-methyl, cyclohexenyl-methyl,piperidinyl-methyl, benzyl, pyridyl-methyl, pyrrolyl-methyl,pyrazolyl-methyl, imidazolyl-methyl, thienyl-methyl, indolyl-methyl,thiazolyl-methyl, furyl-methyl group or COR₄; the optional furthersubstituents being one or more bromine, fluorine, chlorine atom orisopropyl, methyl, phenyl, morpholinyl, or piperidinyl, hydroxy-methylgroup, OR₇, oxo (═O) or SR₇;R₁ and R₂ are independently hydrogen atom or a methyl group, with theproviso that they are not both hydrogen atoms;R₃ is hydrogen or fluorine atom;

R₄ is OR₇,

R₇ is hydrogen atom, an optionally substituted methyl, tert-butyl orbenzyl group, the substituents being one or more fluorine atom, andR₅, R₆, R₈, and R₉ are as defined above,or a pharmaceutically acceptable salt thereof.

Specific compounds of the present invention are listed below:

-   1. tert-butyl    4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate,-   2.    1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,-   3.    2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   4.    (1S)-2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   5.    (1R)-2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   6.    2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   7.    2-[1-(1H-indol-5-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   8.    1-methyl-3-oxo-2-(1-{[1-(propan-2-yl)-1H-indol-3-yl]methyl}piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,-   9.    1-methyl-3-oxo-2-[1-(pyridin-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   10.    2-(1-{[4-(benzyloxy)-1H-indol-3-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   11.    1-methyl-2-{1-[(4-methyl-1,3-thiazol-5-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   12.    2-[1-(2,4-difluorobenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   13.    2-(1-cyclopentylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   14.    1-methyl-2-{1-[(6-methylpyridin-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   15.    2-[1-(furan-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   16.    1-methyl-3-oxo-2-{1-[3-(piperidin-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,-   17.    1-methyl-2-(1-{[6-(morpholin-4-yl)pyridin-2-yl]methyl}piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   18.    2-(1-cyclobutylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   19.    1-methyl-3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   20.    1-methyl-3-oxo-2-[1-(thiophen-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   21.    2-[1-(cyclohexylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   22.    2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   23.    2-{1-[(3-fluoropyridin-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   24.    1-methyl-2-{1-[(1-methyl-1H-pyrrol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   25.    1-methyl-2-{1-[3-(methylsulfanyl)propyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   26.    2-[1-(1H-imidazol-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   27.    2-{1-[(2-chloropyridin-3-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   28.    1-methyl-3-oxo-2-{1-[3-(1H-pyrazol-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,-   29.    1-methyl-2-{1-[(5-methylfuran-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   30.    1-methyl-3-oxo-2-{1-[(3-phenyl-1H-pyrazol-5-yl)methyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,-   31.    2-{1-[(5-bromothiophen-2-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   32.    2-[1-(3-hydroxybenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   33.    2-{1-[2-(difluoromethoxy)benzyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   34.    2-(1-{[5-(hydroxymethyl)furan-2-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   35.    1-methyl-2-[4-(morpholin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   36.    1-methyl-3-oxo-2-[4-(piperidin-1-yl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   37.    1-methyl-2-[4-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   38.    1-methyl-3-oxo-2-[4-(piperidin-1-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   39. tert-butyl    4-(4-carbamoyl-6-fluoro-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate,-   40.    6-fluoro-1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,-   41.    2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   42.    2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   43.    6-fluoro-2-[1-(1H-indol-5-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   44.    6-fluoro-1-methyl-3-oxo-2-[1-(pyridin-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   45.    6-fluoro-1-methyl-2-{1-[(4-methyl-1,3-thiazol-5-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   46.    2-[1-(2,4-difluorobenzyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   47.    2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   48.    6-fluoro-1-methyl-2-{1-[(6-methylpyridin-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   49.    6-fluoro-2-[1-(furan-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   50.    6-fluoro-1-methyl-3-oxo-2-{1-[3-(piperidin-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,-   51.    6-fluoro-1-methyl-2-(1-{[6-(morpholin-4-yl)pyridin-2-yl]methyl}piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   52.    2-(1-cyclobutylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   53.    6-fluoro-1-methyl-3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   54.    6-fluoro-1-methyl-3-oxo-2-[1-(thiophen-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   55.    2-[1-(cyclohexylmethyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   56.    2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   57.    6-fluoro-2-{1-[(3-fluoropyridin-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   58.    6-fluoro-1-methyl-2-{1-[(1-methyl-1H-pyrrol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   59.    6-fluoro-2-[1-(1H-imidazol-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   60.    2-{1-[(2-chloropyridin-3-yl)methyl]piperidin-4-yl}-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   61.    6-fluoro-1-methyl-2-{1-[(5-methylfuran-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   62.    2-{1-[(5-bromothiophen-2-yl)methyl]piperidin-4-yl}-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   63.    6-fluoro-2-(1-{[5-(hydroxymethyl)furan-2-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   64.    (1S)-2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   65.    (1R)-2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   66.    1-methyl-3-oxo-2-[4-(piperidin-4-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   67.    1-methyl-3-oxo-2-[4-(piperazin-1-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   68.    1-methyl-2-{4-[(1-methylpiperidin-4-yl)methyl]phenyl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   69.    1-methyl-2-[3-(1-methylpiperidin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   70.    1-methyl-2-[3-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   71.    2-[4-cyclohexyl-3-(1-methylpiperidin-4-yl)phenyl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   72.    2-[4-cyclohexyl-3-(4-methylpiperazin-1-yl)phenyl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   73.    2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   74.    1-methyl-3-oxo-2-[1-(2,2,2-trifluoroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide    hydrate,-   75.    1-methyl-3-oxo-2-[1-(tetrahydrofuran-3-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   76. tert-butyl    3-{[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]methyl}azetidine-1-carboxylate,-   77.    1-methyl-2-{1-[2-methyl-2-(piperidin-1-yl)propyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   78.    2-{1-[3-(dimethylamino)-2,2-dimethylpropyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   79.    2-{1-[(1-ethyl-4,5-dihydro-1H-pyrazol-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   80.    1-methyl-2-[1-(2-methylbutyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   81.    2-[1-(cyclopentylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   82. methyl    4-[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]butanoate,-   83.    1-methyl-3-oxo-2-[1-(2,2,2-trichloroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   84. ethyl    2-{[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]methyl}cyclopropanecarboxylate,-   85.    2-(1-hexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   86.    1-methyl-2-(1-nonylpiperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   87.    2-[1-(3-cyclohexylpropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   88.    1-methyl-3-oxo-2-[1-(3-phenylpropyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   89.    2-[1-(cyclopropylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   90.    2-(1-heptylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   91.    1-methyl-3-oxo-2-(1-pentylpiperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,-   92.    1-methyl-3-oxo-2-[1-(2-phenylethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   93.    2-[1-(3,3-dimethylbutyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   94.    1-methyl-3-oxo-2-(1-propylpiperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,-   95.    2-[1-(5-hydroxypentyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   96.    1-methyl-3-oxo-2-[1-(2,2,2-trifluoroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,-   97.    2-(1-butylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   98.    1-methyl-2-[1-(2-methylpropyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   99.    1-methyl-2-{1-[(1-methyl-1H-benzimidazol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   100.    2-[1-(2,3-dihydroxypropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   101.    2-[1-(2,2-dimethylpropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   102.    2-[1-(2,2-dimethylpent-4-en-1-yl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   103.    2-(1-ethylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   104.    1-methyl-2-[1-(3-methylbutyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   105.    2-[1-(2-ethylbutyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   106.    2-(1-cyclohexylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   107.    2-(1-cyclopentylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   108.    2-(1-cyclobutylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   109.    2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   110.    2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   111.    2-(1-cyclobutylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   112.    2′-(1-cyclohexylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,-   113.    2′-(1-cyclohexylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,-   114.    2′-(1-cyclopentylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,    115.    2′-(1-cyclopentylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,-   116.    2′-(1-cyclobutylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,-   117.    2′-(1-cyclobutylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1-isoindole]-4′-carboxamide,-   118.    2-(1-cyclohexylpiperidin-4-yl)-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   119.    2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   120.    2-(1-cyclopentylpiperidin-4-yl)-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   121.    2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   122.    2-(1-cyclohexylazetidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   123.    2-(1-cyclohexylazetidin-3-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   124.    2-(1-cyclopentylazetidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   125.    2-(1-cyclopentylazetidin-3-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   126.    2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,-   127.    2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide    and-   128.    2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide    or a pharmaceutically acceptable salt thereof.

The present invention also provides processes for the preparation ofcompounds of formula (I) as defined above. Accordingly, a process of thepresent invention comprises one of the following sequence of steps:

Sequence A)

Step a) Halogenating a Compound of Formula (X):

wherein R₁ and R₃ are as defined above;

Step b) Cyano-De-Aminating the Resultant Compound of Formula (IX):

wherein Hal is halogen such as Cl, Br, I and R₁ and R₃ are as definedabove;

Step c) Hydrolyzing the Resultant Compound of Formula (VIII):

wherein Hal, R₁ and R₃ are as defined above;

Step d) Hydrolyzing the Resultant Compound of Formula (VII):

wherein Hal, R₁ and R₃ are as defined above;

Step e) Esterifying the Resultant Compound of Formula (VI):

wherein Hal, R₁ and R₃ are as defined above;

Step f) Cyclizing the Resultant Compound of Formula (V):

wherein T is C₁-C₆ alkyl or aryl C₁-C₆ alkyl group and R₁ and R₃ are asdefined above with a suitable amine of formula (XIV):

H₂N—R  XIV

wherein R is as defined above;

Step g) Alkylating the Resultant Compound of Formula (IV):

wherein Hal, R, R₁ and R₃ are as defined above;

Step h) Cyano-De-Halogenating the Resultant Compound of Formula (III):

wherein Hal, R, R₁, R₂ and R₃ are as defined above;

Step i) Hydrolyzing a Compound of Formula (II):

wherein R, R₁, R₂ and R₃ are as defined above to give a compound offormula (I):

wherein R, R₁, R₂ and R₃ are as defined above; or

Sequence B):

Step l) Halogenating a Compound of Formula (XI):

wherein R₁ and R₃ are as defined above;

Step e and m) Esterifying the Resultant Compound of Formula (VI) asDefined Above and Cyano-De-Halogenating the Resultant Compound ofFormula (V) as Defined Above;

Step f′) Cyclizing the Resultant Compound of Formula (XII)

wherein T, R₁ and R₃ are as defined above, with a suitable amine offormula (XIV) as defined above;

Step g′) Alkylating the Resultant Compound of Formula (XIII):

wherein R, R₁ and R₃ are as defined above;

Step i′) Hydrolyzing the Resultant Compound of Formula (XIII) as DefinedAbove to Give a Compound of Formula (I) as Defined Above, or

Sequence C)

either Step n) a reductive amination of a compound of formula (XV):

wherein R₁ and R₃ are as defined above with a suitable amine of formula(XIV) as defined above;or Step q) a reductive alkylation of a compound of formula (XX):

wherein R₁ and R₃ are as defined above;

Step o) Reacting with Maleic Anhydride Under Diels-Alder Conditions theResultant Compound of Formula (XVI):

wherein R, R₁ and R₃ are as defined above;

Step p) Aromatizing the Resultant Compound of Formula (XVII):

wherein R, R₁ and R₃ are as defined above andeither Step i″) amidating the resultant compound of formula (XVIII):

-   -   wherein R, R₁ and R₃ are as defined above and R₂ is hydrogen        atom so as to obtain a compound of formula (I) as defined above        wherein R₂ is hydrogen atom, or

Step e′) Esterifying a Compound of Formula (XVIII) as Defined AboveWherein R is as Defined Above;

Step s) N-Deprotecting the Resultant Compound of Formula (XXI):

wherein T, R₁ and R₃ are as defined above and R is as defined above;

Step t) N-Alkylating the Resultant Compound of Formula (XXII):

wherein T, R₁ and R₃ are as defined above and;either Step r) hydrolyzing the resultant compound of formula (XXI) asdefined above wherein R is as defined above;

-   or Step g″) alkylating a compound of formula (XXI) as defined above    wherein R is as defined above;    -   Step r′) hydrolyzing the resultant compound of formula (XXIII):

wherein T, R₁, and R₃ are as defined above and R₂ is not hydrogen atomand then converting the resultant compound of formula (XVIII) wherein R,R₁ and R₃ are as defined above and R₂ is not hydrogen atom operating asdescribed above under step i″).

If necessary or wanted, the processes above described comprisesconverting a compound of formula (I) into a different compound offormula (I) by known chemical reactions; and/or, if desired, convertinga compound of formula (I) into a pharmaceutically acceptable saltthereof or converting a salt into a free compound of formula (I).

Also an intermediate compound of formula (II), (III) and (XVIII) asdefined above may be converted into a different compound of formula(II), (III) and (XVIII).

The known chemical reactions for possible conversions of compounds ofthe formula (I), (II), (III) and (XVIII) as defined above into adifferent compound of the formula (I), (II), (III) and (XVIII) asdefined above are for example:

1Cv) an alkylation of a compound of formula (II) or (III) as definedabove;2Cv) a protection of a compound of formula (XVIII) as defined above;3Cv) a deprotection of a compound of formula (I) or (II) as definedabove;4Cv) a reductive amination of a compound of formula (I) or (II) asdefined above.

All the above processes are analogy processes which can be carried outaccording to well known methods and under suitable conditions known inthe art.

The synthesis of a compound of formula (I), according to the syntheticprocesses described above, can be carried out in a stepwise manner,whereby each intermediate is isolated and purified by standardpurification techniques, like, for example, column chromatography,before carrying out the subsequent reaction. Alternatively, two or moresteps of the synthetic sequence can be carried out in a so-called“one-pot” procedure, as known in the art, whereby only the compoundresulting from the two or more steps is isolated and purified.

Schemes 1-2 below show the preparation of a compound of formula (I) asdefined above.

According to step a), a compound of formula (IX) can be obtained byhalogenating a compound of formula (X) in different ways andexperimental conditions known in the art. Preferably this reaction iscarried out in the presence of N-bromosuccinimide, N-iodosuccinimmide,N-chlorosuccinimide, bromine, iodine, hydrobromic acid/hydrogenperoxide, in a suitable solvent, such as acetonitrile,N,N-dimethylformamide, dioxane, dimethylsulfoxide, acetic acid, water ata temperature ranging from about room temperature to reflux and for aperiod of time ranging from about 1 hour to about 96 hours.

According to step b), a compound of formula (VIII) can be obtained by atwo-reactions sequence from a compound of formula (IX) in different waysand experimental conditions known in the art. First reaction ispreferably carried out in the presence of sodium nitrite/hydrochloricacid or tert-butylnitrite in a suitable solvent, such astetrahydrofurane, dimethoxyethane, dimethylsulfoxide, acetic acid, waterat a temperature ranging from about −20° C. to room temperature and fora period of time ranging from 10 minutes to about 24 hours. Second stepis preferably carried out in the presence of sodium, copper or potassiumcyanide, often in the presence of an additive such as copper orpotassium chloride, in a suitable solvent such as tetrahydrofuran,dimethoxyethane, dimethylsulfoxide, acetic acid, toluene, water at atemperature ranging from about −20° C. to reflux and for a period oftime ranging from 10 minutes to about 96 hours. According to step c),the hydrolysis of a compound of formula (VIII) to give a compound offormula (VII) can be carried out in different ways, according toconventional methods for transforming a cyano group to amide. Preferablythis reaction is carried out in a suitable solvent such as, forinstance, methanol, ethanol, butanol, 1,4-dioxane, water, or a mixturethereof, in the presence of a suitable acid or base, such as, forinstance, sulfuric acid, hydrochloric acid, sodium hydroxide, sodiumcarbonate, or a suitable reagent such as hydrogen peroxide or sodiumperborate. Typically, the reaction is carried out at a temperatureranging from room temperature to reflux and for a time ranging fromabout 1 hour to about 96 hours.

According to step d) a compound of formula (VII) can be converted into acompound of formula (VI) according to conventional methods. Preferablythe reaction is carried out in the presence of water by treatment with abase such as potassium or sodium carbonate, potassium or sodiumhydroxide, in a suitable solvent such as, for instance, methanol,ethanol, at a temperature ranging from room temperature to reflux, for atime ranging from about 30 min. to about 96 hours. Alternatively thisreaction can be carried out in the presence of sodium nitrite/aceticacid, sulfuric acid, fosforic acid at a temperature ranging from roomtemperature to reflux and for a time ranging from about 1 hour to about96 hours.

According to step e) a compound of formula (VI) can be converted into acompound of formula (V) according to conventional methods. Preferablythe reaction is carried out in the presence of hydrochloric acid,sulfuric acid, acetic acid in a solvent like methanol, ethanol, water,or a mixture thereof, at a temperature ranging from room temperature toreflux and for a time ranging from about 1 hour to about 96 hours.Alternatively, this reaction can be carried out with alkyl iodide orbromide in the presence of a suitable base such as sodium or potassiumcarbonate, sodium, lithium or potassium hydroxide at a temperatureranging from room temperature to reflux and for a time ranging fromabout 1 hour to about 96 hours.

According to step f), a compound of formula (IV) can be obtained by atwo-reactions sequence from a compound of formula (V) in the presence ofa compound of formula (XIV) in different ways and experimentalconditions known in the art. First reaction is preferably carried out inthe presence of N-bromosuccinimide with a radical initiator such asbenzoyl peroxide or azobisisobutyronitrile in a suitable solvent, suchas carbon tetrachloride, chloroform, dichloromethane or methyl pivalateat a temperature ranging from about room temperature to reflux and for aperiod of time ranging from 10 minutes to about 24 hours. Secondreaction can be carried out both under basic or acidic conditions suchas in the presence of sodium or potassium carbonate,1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine,N,N—N,N-diisopropylethylamine, pyridine or acetic acid, hydrochloricacid in a suitable solvent, such as tetrahydrofurane, dimethoxyethane,1,4-dioxane, toluene at a temperature ranging from room temperature toreflux and for a period of time ranging from 1 hour to about 96 hours.

According to step g) the conversion of a compound of formula (IV) into acompound of formula (III) can be carried out in different ways,according to conventional methods for C-alkylation reactions.Preferably, this reaction is carried out in a suitable solvent such as,for instance, tetrahydrofuran, diethyl ether, dioxane,N,N-dimethylformamide, dimethoxyethane, in the presence of a suitablebase, such as, for instance, lithium, sodium or potassiumbis(trimethylsilyl)amide, sodium, potassium or cesium carbonate, sodiumhydride, at a temperature ranging from −78° C. to room temperature andfor a time ranging from about 10 minutes to about 24 hours. Alkylatingagent is usually a halogen or a sulphonates derivative; most often theleaving group is iodo, bromo, triflate or mesylate.

According to step h), the conversion of a compound of formula (III) intoa compound of formula (II) can be carried out in different ways,according to conventional methods for cyanation reactions. Preferably,this reaction is carried out in the presence of copper cyanide orpotassium hexacyanoferrate (II) as cyano source in a suitable solventsuch as, for instance, methanol, ethanol, tetrahydrofuran, 1,4-dioxane,toluene, xylene, N-methyl-2-pyrrolidone, or a mixture thereof, at atemperature ranging from room temperature to reflux and for a timeranging from about 1 hour to about 96 hours. If a catalyst is required,it is usually a metal, most often a palladium derivative such as, forinstance, palladium(II) chloride or palladium(II) acetate in thepresence of a suitable base such as, for instance, sodium, potassium orcesium carbonate, cesium fluoride.

According to step i) the conversion of a compound of formula (II) into acompound of formula (I) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step c).

According to step l), the halogenation of a compound of formula (XI)into a compound of formula (VI) can be carried out in different ways,according to conventional methods for halogenation reactions.Preferably, this reaction is carried out with tetrabutylammonium bromideand/or iodine in the presence of phenyliodine(III) bis(trifluoracetate)or phenyliodo(III) diacetate as halogen source in a suitable solventsuch as, for instance, N,N-dimethylformamide or dichloroethane, at atemperature ranging from room temperature to reflux and for a timeranging from about 1 hour to about 48 hours. The catalyst is usually ametal, most often a palladium derivative such as, for instance,palladium(II) chloride or palladium(II) acetate.

According to step m) the cyanation of a compound of formula (V) to givea compound of formula (XII) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step h).

According to step f) the transformation of a compound of formula (XII)into a compound of formula (XIII) in the presence of a compound offormula (XIV) can be carried out in different ways and experimentalconditions. Preferably it is carried out in a way analogous to thatreported for step f).

According to step h′) the cyanation of a compound of formula (IV) togive a compound of formula (XIII) can be carried out in different waysand experimental conditions. Preferably it is carried out in a wayanalogous to that reported for step h).

According to step g′) the transformation of a compound of formula (XIII)into a compound of formula (II) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step g).

According to step i) the conversion of a compound of formula (XIII) intoa compound of formula (I) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step c).

According to step n) the reductive amination of a compound of formula(XV) to give a compound of formula (XVI), by reaction with a suitableamine of formula (XIV), can be carried out in different ways, accordingto conventional methods for carrying out reductive amination.Preferably, this reaction is carried out in a suitable solvent such as,for instance, methanol, N,N-dimethylformamide, dichloromethane,tetrahydrofuran, benzene, toluene, or a mixture thereof, in the presenceof a suitable reducing agent such as, for instance, sodium borohydride,tetraalkylammonium borohydride, sodium cyanoborohydride, sodiumtriacetoxyborohydride, tetramethylammonium triacetoxy borohydride and inpresence of an acid or basic catalyst, such as, for instance, aceticacid, trifluoroacetic acid, zinc chloride, zinc bromide, tin(IV)chloride, titanium(IV) chloride, boron trifluoride or triethylamine,N,N—N,N-diisopropylethylamine, pyridine at a temperature ranging fromabout 0° C. to reflux and for a time ranging from about 1 hour to about96 hours.

According to step o) the Diels-Alder reaction of a compound of formula(XVI) to give a compound of formula (XVII) can be carried out indifferent ways, according to conventional methods for carrying out thesereactions. Preferably, this reaction is carried out in a suitablesolvent such as, for instance, tetrahydrofuran, benzene, toluene,o-xilene in the presence of maleic anhydride at a temperature rangingfrom about room temperature to reflux and for a time ranging from about1 hour to about 96 hours.

According to step p) the conversion of a compound of formula (XVII) intoa compound of formula (XVIII) can be carried out in different ways,according to conventional methods. Preferably, this reaction is carriedout in a suitable solvent such as, for instance, tetrahydrofuran,toluene, water, in the presence of hydrochloric acid, p-tolensulfonicacid, phosphoric acid, at a temperature ranging from about roomtemperature to reflux and for a time ranging from about 1 hour to about24 hours.

According to step i″) a compound of formula (I) can be obtained byreacting a compound of formula (XVIII) in different ways andexperimental conditions, which are widely known in the art ofcondensation reactions. Preferably a compound of formula (XVIII) isreacted with ammonia or ammonia source such as ammonium salts, in thepresence of an activating agent such as carbonyldiimidazole,benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate,2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate),(O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, dicyclohexyl carbodiimide, diisopropylcarbodiimide, 1-ethyl-3-(3′-dimethylamino) carbodiimide hydrochloricacid salt, optionally in the presence of hydroxybenzotriazole.Preferably, this reaction is carried out in a suitable solvent such as,for instance, N,N-dimethylformamide, N,N-dimethylacetamide,tetrahydrofuran, dichloromethane, 1,4-dioxane, and in the presence of aproton scavenger such as, for example, pyridine, triethylamine,N,N-diisopropylethylamine, at a temperature ranging from roomtemperature to reflux, for a time ranging from about 30 min. to about 96hours.

According to step q) the conversion of a compound of formula (XX) into acompound of formula (XVI) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step n).

According to step e′) the conversion of a compound of formula (XVIII)into a compound of formula (XXI) can be carried out in different waysand experimental conditions. Preferably it is carried out in a wayanalogous to that reported for step e).

According to step r) a compound of formula (XXI) can be transformed intoa compound of formula (XVIII) according to conventional methods.Preferably the reaction is carried out in the presence of water bytreatment with a base such as potassium or sodium carbonate, potassiumor sodium hydroxide, in a suitable solvent such as, for instance,methanol, ethanol, at a temperature ranging from room temperature toreflux, for a time ranging from about 30 min. to about 96 hours.Alternatively this reaction can be carried out in the presence ofhydrochloric acid, sulfuric acid, phosphoric acid at a temperatureranging from room temperature to reflux and for a time ranging fromabout 1 hour to about 96 hours.

According to step s), a compound of formula (XXI) can be transformedinto a compound of formula (XXII) by deprotection of the nitrogen atomaccording to conventional methods enabling the selective hydrolysis oftert-butoxycarbonyl, benzyl, 4-methoxybenzyl, 2,4-dimethoxybenzyl andtriphenylmethyl, phenylethyl protective groups. Preferably this reactionis run under acidic conditions, for instance in the presence of aninorganic or organic acid such as hydrochloric, trifluoroacetic ormethanesulfonic acid, in a suitable solvent such as dichloromethane,1,4-dioxane, a lower alcohol, such as methanol or ethanol, at atemperature ranging from room temperature to reflux and for a period oftime ranging from about 1 hour to about 48 hours. In alternative, thisreaction is carried out under reducing condition, such as, for instance,in the presence of molecular hydrogen and a hydrogenation catalyst in asuitable solvent such as ethanol, methanol, ethyl acetate, or a mixturethereof. The catalyst is usually a metal, most often a palladiumderivative such as, for instance, palladium on carbon, palladiumhydroxide or palladium black. Alternatively, the removal of protectivegroup base-labile can be carried out by using a base such as potassium,sodium or cesium carbonate, potassium or sodium hydroxide, pyridine,triethylamine, N,N-diisopropylethylamine, in a suitable solvent such asmethanol, ethanol, at a temperature ranging from room temperature toreflux and for a period of time ranging from about 1 hour to about 48hours.

According to step t), the conversion of a compound of formula (XXII)into a compound of formula (XXI) can be carried out in different ways,according to conventional methods for N-alkylation reactions.Preferably, this reaction is carried out in a suitable solvent such as,for instance, tetrahydrofuran, dioxane, N,N-dimethylformamide,dimethoxyethane, dimethylsulfoxide, benzene in the presence of asuitable base, such as, for instance, triethylamine,N,N-diisopropylethylamine, sodium, potassium or cesium carbonate, sodiumhydride, sodium methoxide, lithium, sodium or potassiumbis(trimethylsilyl)amide at a temperature ranging from −78° C. to refluxand for a time ranging from about 1 hour to about 96 hours. Alkylatingagent is usually a halogen or a sulphonates derivative; most often theleaving group is iodo, bromo, triflate or mesylate.

According to step g″) the conversion of a compound of formula (XXI) intoa compound of formula (XXIII) can be carried out in different ways andexperimental conditions. Preferably it is carried out in a way analogousto that reported for step g).

According to step r′) the conversion of a compound of formula (XXIII)into a compound of formula (XVIII) can be carried out in different waysand experimental conditions. Preferably it is carried out in a wayanalogous to that reported for step r).

According to the conversion described under 1Cv) the alkylation of acompound of formula (II) or (III) can be carried out in different waysand experimental conditions. Preferably it is carried out in a wayanalogous to that reported for step g).

According to the conversion described under 2Cv), the reaction of acompound of formula (XVIII) may be carried out in different ways andexperimental conditions. Preferably when the protective group istert-butoxycarbonyl, the reaction may be carried out in the presence ofdi-tert-butyl dicarbonate in different solvents such as methanol,ethanol, acetonitrile, tetrahydrofurane, dichloromethane in the presenceof a base such as pyridine, N,N-dimethylaminopyridine, triethylamine,sodium or potassium carbonate at a temperature ranging from roomtemperature to reflux and for a time ranging from about 1 hour to about96 hours.

According to the conversion described under 3Cv), the deprotection of acompound of formula (I) or (II) may be carried out in different ways andexperimental conditions. Preferably when the protective group istert-butoxycarbonyl, the reaction may be carried out in the presence ofhydrochloric acid, trifluoroacetic acid in a suitable solent such asdioxane, dichloromethane, tetrahydrofurane at a temperature ranging fromroom temperature to reflux and for a time ranging from about 1 hour toabout 24 hours.

According to the conversion described under 4Cv), the reductiveammination of a compound of formula (I) or (II) may be carried out indifferent ways and experimental conditions. Preferably it is carried outin a way analogous to that reported for step n).

Substituted isoindoles can be prepared using standard procedures inorganic synthesis as reported, for instance, in Smith, Michael—March'sAdvanced Organic Chemistry: reactions mechanisms and structure—6thEdition, Michael B. Smith and Jerry March, John Wiley & Sons Inc., NewYork (NY), 2007. It is known to the skilled person that conversion of achemical function into another may require that one or more reactivecenters in the compound containing this function be protected in orderto avoid undesired side reactions. Protection of such reactive centers,and subsequent deprotection at the end of the synthetic conversions, canbe accomplished following standard procedures described, for instance,in: Green, Theodora W. and Wuts, Peter G. M. —Protective Groups inOrganic Synthesis, Third Edition, John Wiley & Sons Inc., New York (NY),1999.

In cases where a compound of formula (I) contains one or more asymmetriccenters, said compound can be separated into the single isomers byprocedures known to those skilled in the art. Such procedures comprisestandard chromatographic techniques, including chromatography using achiral stationary phase, or crystallization. General methods forseparation of compounds containing one or more asymmetric centers arereported, for instance, in Jacques, Jean; Collet, André; Wilen, SamuelH.,—Enantiomers, Racemates, and Resolutions, John Wiley & Sons Inc., NewYork (NY), 1981.

As stated above, a compound of formula (I) can be converted into apharmaceutically acceptable salt according to standard procedures thatare known to those skilled in the art. Alternatively, a compound offormula (I) that is obtained as a salt can be converted into thecorresponding free base or the free acid according to standardprocedures that are known to the skilled person.

The starting materials of the process of the present invention, i.e.compounds of formula (X), (XI), (XIV), (XV) and (XX) are eithercommercially available or can be prepared by using well-known methods.

For example, the compound of formula (XX) can be easily obtainedaccording to conventional procedures, which are widely known in the artfor reductive amination reactions (see for example Demir, Ayhan et al.Helvetica Chimica Acta (2003), 86(1), 91-105).

The intermediate compounds of formula (II), (III), (IV), (XIII),(XVIII), (XXI), (XXII) and (XXIII) are novel and are also within thescope of the present invention.

Biochemical Assay

Affinity evaluation of the tested compounds and their selectivity withrespect to the different PARP isoforms of interest was assessed in adisplacement assay.

The identification of compounds capable of binding several PARP proteinsis carried out through a screening method including the steps of

a) providing a reaction mixture containing:the PARP protein isoform under investigation,a compound of formula (IP):

wherein R₁₁ is hydrogen atom or a methyl group, B is (CH₂)_(n)—NH groupwherein n is 2 to 6; m is 0 or 1 and X⁻⁻⁻ is a counterion, andserial dilutions of the test compound;b) comparing the polarization signal generated in the absence of thetest compound with the one generated in the presence of differentconcentrations of the test compound, andc) evaluating the ability of the test compound to displace the compoundof formula (IP) as defined above indicated from a decreased fluorescencepolarization level.

Preferably, for the screening method above cited, either the PARPprotein and the 5H-Phenanthridin-6-one-derived probe of formula (IP) arepre-mixed, or the PARP protein and the test compound are pre-mixed. In afurther preferred screening method, the PARP proteins are PARP-1, PARP-2and PARP-3. The term “PARP protein” encompasses full-length nativeproteins as well as fragment thereof.

More preferably, R₁₁ is hydrogen or methyl, m is 0 or 1; when m is 1, nis 3 or 6, X⁻⁻⁻ is trifluoroacetate.

The 5H-phenanthridin-6-one-derived probe (IP) was selected for itscapability of binding to the PARP proteins, both encompassingfull-length native proteins and fragment thereof.

The polarization signal can be measured, e.g., by a plate reader such asthe Saphire2 (Tecan). The displacement ability of the test compound isin correlation with the compound affinity for the NAD pocket of theenzyme. Affinity binding constant (KD) and DC₅₀s of the test compoundcan be determined as explained in the Example section. The assay usedfor testing the compounds of the invention is based on the use of knownPARP inhibitors chemically conjugated to a moiety that provides thefluorescent signal. Specific compounds used are:

-   P1.    9-Dimethylamino-11,11-dimethyl-1-(3-{methyl-[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-carbamoyl}-propyl)-2,3,4,11-tetrahydro-naphtho[2,3-g]quinolinium    trifluoroacetate;-   P2.    9-Dimethylamino-11,11-dimethyl-1-[3-(3-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-propylcarbamoyl)-propyl]-2,3,4,11-tetrahydro-naphtho[2,3-g]quinolinium    trifluoroacetate;-   P3.    9-Dimethylamino-11,11-dimethyl-1-[3-(6-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-hexylcarbamoyl)-propyl]-2,3,4,11-tetrahydro-naphtho[2,3-g]quinolinium    trifluoroacetate.

A compound of formula (IP) as defined above can be prepared through aprocess comprising:

step u) either reacting a compound of formula (XXIV):

wherein Lg is a leaving group, preferably a chlorine atom, with acompound of formula (XXV): R₁₁—NH₂ wherein R₁₁ is as defined above, togive a compound of formula (XXVI):

wherein R₁₁ is as defined above and m is 0;or step v) reacting a compound of formula (XXIV) as defined above with acompound of formula XXVa:

wherein R₁₁ and B are as defined above, m is 1 and R₁₂ is hydrogen atom,methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl,9-fluorenylmethoxycarbonyl or the like; and

step z) converting if necessary the resultant compound of formula(XXVIa):

wherein R₁₁ and B are as defined above, m is 1 and R₁₂ is amethoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl,9-fluorenylmethoxycarbonyl group or the like, into a compound of formula(XXVI) as defined above;

step ii) coupling the resultant compound of formula (XXVI) as definedabove with a compound of formula (XXVII):

wherein X⁻ is as defined above so as to obtain the final compound offormula (IP) as defined above; and, if necessary or wanted, converting acompound of formula (IP) into another compound of formula (IP) whereinX⁻ is different.

The starting compound of formula (XXIV) can be prepared as extensivelydescribed, see, for instance WO 2001042219; the compounds of formula(XXVII) are described in Cha, J. H. et al., J. Med. Chem. 2005, 48,7513-7516 and ATTO 610 fluorescent moiety activated as its NHS ester iscommercially available (Naphtho[2,3-g]quinolinium,9-(dimethylamino)-1-[4-[(2,5-dioxo-1-pyrrolidinyl)oxy]-4-oxobutyl]-2,3,4,11-tetrahydro-11,11-dimethyl-,perchlorate (1:1), ATTO-TEC GmbH, Siegen, Germany).

Scheme 3 below shows the preparation of compound of formula (IP) whereinR₁₁, R₁₂, B and m have the meanings defined above.

According to steps u and v of the process, a compound of formula (XXIV)as defined above is reacted with a compound of formula (XXV) or (XXVa)as defined above, in the presence of a base such as, for instance,sodium or potassium hydroxide, sodium, potassium or cesium carbonate,sodium or potassium hydrogencarbonate, triethylamine,N,N-diisopropylethylamine, pyridine and the like, in a suitable solventsuch as acetonitrile, dioxane, methanol, ethanol orN,N-dimethylformamide at a temperature ranging from 0° C. to reflux togive, starting from compound (XXV), as defined above, a compound of theformula (XXVI), as defined above or, starting from compound (XXVa), asdefined above, a compound of formula (XXVIa), as defined above;

According to step z of the process, a compound of formula (XXVIa),wherein m=1, R₁₂ is a methoxycarbonyl, ethoxycarbonyl,tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl groupor the like, is converted into a compound of formula (XXVI), as definedabove, by removing the corresponding nitrogen protective group. Inparticular, protective groups such as tert-butoxycarbonyl and the likecan be removed under acidic conditions, preferably in the presence of aninorganic or organic, acid such as hydrochloric, trifluoroacetic ormethanesulphonic acid in a suitable solvent such as dichloromethane,dichloroethane, dioxane, a lower alcohol, such as methanol or ethanol,at a temperature ranging from room temperature to reflux. Protectivegroups such as benzyloxycarbonyl and the like can be removed with asuitable reducing agent, such as molecular hydrogen, cyclohexene,cyclohexadiene, formic acid, ammonium formate and the like, in thepresence of a hydrogenation catalyst, such as, for instance, palladiumon carbon, palladium hydroxide, palladium black, Ni Raney and the like,in a suitable solvent, such as methanol, ethanol, dioxane and the likeat a temperature ranging from room temperature to reflux.

Protective groups such as methoxycarbonyl, ethoxycarbonyl,9-fluorenylmethoxycarbonyl and the like could be removed under basicconditions such as, for instance, sodium, potassium or cesium carbonate,sodium, potassium or barium hydroxide, hydrazine, piperidine, morpholineand the like in a suitable solvent such as methanol, ethanol, water,N,N-dimethylformamide, N,N-dimethylacetamide and the like, at atemperature ranging from room temperature to reflux.

According to step ii of the process, a compound of formula (XXVI), asdefined above, is reacted with a compound of formula (XXVII) in thepresence of a suitable base such as triethylamine,N,N-diisopropylethylamine, pyridine and the like, in a solvent such asN,N-dimethylformamide, N,N-dimethylacetamide and the like, at atemperature ranging from 0° C. to room temperature so as to obtain thedesired compounds of formula (IP).

The assay is based on the use of a probe of formula (IP) that binds tothe NAD binding pocket and takes advantage of the significant change inthe polarization signal observed upon binding of the probe to PARP-1, -2and -3.

The probe of formula (IP) was tested for its ability to bind FL PARP-1,-2 and -3 in a titration experiment. The assay performances were thenevaluated (Z′ factor) as well as the displacement of the probe by itsscaffold and known commercially available PARP inhibitors. In all of theexperiments, the polarization signal was measured using a Saphire2 platereader (Tecan). Data analysis was performed using Dynafit software. Inparticular, titration data were fitted to the following equilibria:Enzyme+probe<==>Complex Enzyme-probe, while displacement data werefitted to the following equilibria: Enzyme+probe<==>ComplexEnzyme-probe, Enzyme+Compound<==>Complex Enzyme-Compound, wherebybinding of probe and compound on the enzyme are mutually exclusive (purecompetitive mechanism). Displacement data were also fitted using Excellspreadsheet (Microsoft Inc. Seattle, USA) to a four parameter logisticmodel (4PL), or Hill-Slope model to calculate DC₅₀s where the DC₅₀ valuerepresents the compound concentration at which the polarization signalis diminished by 50% compared to untreated controls.

The titration experiment was performed as follows: 50 nM probe (compoundP1), FL PARP-1, -2 and -3 at concentrations from 5 μM to 0, withdilution steps 1:1.5 in 50 mM TrisHCl, pH 7.8, 150 mM sodium chloride,10 mM magnesium chloride, 0.001% Triton X100, 1% DMSO (buffer 1). Asimilar procedure was followed for compound P3 titration.

The obtained results (shown in Table 1 below) indicated that the probe(compound P1) is capable to bind all of the tested isoforms of PARP. Forcompound P3 only PARP-1 KD is reported. The Z′ factor(Z′=1-(3*(SDprobe+protein+SDprobe)/(Meanprobe+protein−Meanprobe)) wasdetermined as follows: 50 nM probe (compound P1), 250 nM of PARP-1 and-2, 200 nM of PARP-3. PARP-1 concentration was equal to 100 nM whencompound P3 was used as probe. In all cases, the Z's were higher than0.7 indicating that the assays were robust (table 1).

TABLE 1 KD (μM) STD (μM) Z′ PARP-1 FL (compound P3) 0.4 0.07 0.75 PARP-1FL (compound P1) 1.04 0.14 0.73 PARP-2 FL (compound P1) 1.05 0.2 0.78PARP-3 FL (compound P1) 0.18 0.016 0.9

The assay was validated using 3-aminobenzamide (3-AB) and PJ-34(N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-N,N-dimethylacetamide) in adisplacement assay performed as follows: serial dilutions of testcompounds were first prepared in 100% DMSO and further diluted in assaybuffer 1 in order to have a 1% final DMSO concentration. 3-AB weretested at 100 μM as highest concentration, while 10 μM was PJ-34 highestconcentration. The enzymes were present at a final concentration of 250nM for PARP-1 (100 nM when compound P3 was used as probe) and PARP-2while 200 nM was used for PARP-3. Probe (compound P1 or compound P3)final concentration was 50 nM. The mixture of enzyme and probe (compoundP1 or compound P3) was added to the previously diluted compounds.Results (Table 2) indicate that the probes (compound P1 or compound P3)could be fully displaced by 3-AB and PJ-34 from all of the tested PARPisoforms indicating that the probes (compound P1 or compound P3) bindingis specific. In agreement, affinity binding constants (KD) weredetermined by fitting with a pure competitive mechanism. KD values, arethe average of three independent experiments.

3-AB, as expected, was not selective among the PARP isoforms and showeda lower affinity with respect to PJ-34.

TABLE 2 PJ34 KD STD 3-AB KD STD (μM) (μM) (μM) (μM) PARP-1 FL (compoundP3) <0.01* — 5.56 0.55 PARP-1 FL (compound P1) <0.03* — 6.68 1.2 PARP-2FL (compound P1) <0.03* — 7.4 1.04 PARP-3 FL (compound P1) 0.15 0.02617.7 4.25 *assay sensitivity limits based on a fitting error <50%

Taken together, these results show that the displacement assay isspecific. Moreover it allows quantitative potency evaluation of standardPARP inhibitors tested, and therefore selectivity evaluation amongassays. The same assay, by using either probe P1 or P3, was used toevaluate compounds of formula (I) as reported in table 3.

TABLE 3 PARP-1 PARP-2 PARP-3 Compound (DC₅₀μM) (Kd μM)* (DC₅₀μM)(DC₅₀μM) 3 <0.25 <0.01 >10 >10 4 <0.25 <0.01 >10 >10 5 2.11 0.78 >10 >106 <0.25 0.03 >10 >10 7 <0.25 0.03 >10 0.52 8 0.44 0.25 >10 3.1 9 0.350.17 >10 >10 13 <0.25 0.05 1.46 >10 15 <0.25 0.04 >10 >10 18 <0.25 <0.011.82 >10 19 <0.25 0.02 >10 3.04 20 <0.25 <0.01 >10 3.86 21 <0.250.03 >10 >10 22 <0.25 0.04 >10 >10 26 <0.25 0.07 6.88 2.83 35 <0.250.04 >10 >10 36 <0.25 0.08 >10 >10 38 1.11 — >10 >10 126 <0.25 — >10 >10127 0.41 0.06 >10 >10 128 5.66 0.87 >10 >10 *Assay performed withcompound P3 as the probe. In all other cases compound P1 was used as theprobe. From the above data, it is clear to a person skilled in the artthat compounds of formula (I) of the present invention are highly potentas PARP-1 inhibitors and extremely selective versus PARP-2 and PARP-3(compare PARP-1 Kd, PARP-2 DC₅₀ and PARP-3 DC₅₀ values).

Pharmacology

PARP-1 is a DNA damage-induced polymerase that catalyzes the cleavage ofNAD+ into nicotinamide and ADP-ribose and then uses the latter tosynthesize branched nucleic-acid like polymers poly(ADP-ribose). Invivo, the most abundantly poly (ADP-ribosylated) protein is PARP-1itself, followed by histones. PARP-1 is responsible for 90% of this DNAdamage-induced activity while the remaining 10% is due to PARP-2.

Cellular Assay

Cellular activity of PARP-1 inhibitors was assessed by measuring theinhibition of the hydrogen peroxide induced PAR formation in HeLa cells(ECACC). Cellular PAR levels were measured by immunocytochemistry, andquantified using an ArrayScan vTi instrument (Cellomics ThermoScientific).

Studies were performed as follows: 6000 cells/well were seeded in 96well plates (Perkin Elmer) in MEM/10% FCS and incubated for 24 hours at37° C., 5% carbon dioxide. Test compounds were then added at therequired concentration for 30′. DNA damage was then induced addinghydrogen peroxide at the concentration of 0.1 mM for 15 min.Concentration curves were prepared in MEM/10% FCS from compound stocksin DMSO, and final DMSO concentration was 0.002% (v/v). Duplicate wellsfor each concentration point were prepared with a typical highestcompound concentration of 20 μM and serial dilution 1:3. Plates weredried and fixed adding cold methanol-acetone (70:30) solution for 15 minat room temperature, fixing solution was aspired and wells were airdried for 5 min and then dehydrated in PBS. Non-specific binding siteswere blocked by incubating wells for 30 min in PBS containing 5% (w/v)FBS 0.05% tween20. Wells were then incubated for 1 hour at roomtemperature in PBS containing anti PAR mouse monoclonal antibody(Anti-PAR, Mouse mAb 10H, Tulip Cat N^(o) 1020) diluted 1:200 inblocking solution. After 3 washes in PBS, wells incubated in PBS (w/v)5% FBS 0.05% Tween20 containing 2 μg/mL Cy2-conjugated Goat anti mousesecondary antibody (Amersham Pharmacia Biotech cat. N^(o) PA 42002)(Absorption maximum 489 nm fluorescence maximum 506 nm) and 1 μg/mL DAPI(Absorption maximum 359 nm fluorescence maximum 461 nm)(4′,6-Diamidino-2-phenyindole dilactate) (Sigma cat. N^(o) D9564), ahigh sensitivity dye for nucleic acid staining. After washing further 3times in PBS, cellular PAR immunoreactivity was assessed using theArrayScan vTi instrument, with a Zeiss 10×0.5 N.A. objective, andapplying the Cytotoxicity.V3 algorithm (Cellomics/Thermo Fisher) with aXF100 filter. At least 10 fields, corresponding to at least 900 cells,were read for each well. IC₅₀ values represent the compoundconcentration at which cellular PAR signal is diminished by 50% comparedwith untreated controls.

The following formula is used:

IC ₅₀=Bottom+(Top−Bottom)/(1+10̂((Log EC ₅₀ −X)));

X is the logarithm of concentration. IC₅₀ is the response; IC₅₀ startsat Bottom and goes to Top with a sigmoid shape. Given the above assays,compounds of formula (I) of the present invention inhibited PARformation with IC₅₀ values lower than 10 μM, as depicted in table 4below.

TABLE 4 PAR assay Compound (IC₅₀ μM) 3 0.28 4 0.15 5 6.3 6 1.3 7 7 9 313 0.5 18 2.7 19 2.5 21 1.2 35 4.9 36 9 38 5.3 126 0.27 127 0.64 128 4

Pharmacokinetics

The pharmacokinetic profile and the oral bioavailability of thecompounds have been investigated in the mouse (Balb, Nu/Nu, Harlan,Italy) in ad hoc pharmacokinetic studies. The compounds were formulatedin 10% tween 80/dextrose for intravenous bolus administration while oraladministrations were performed using the compounds formulated in 0.5%methylcellulose. A single administration at the dose of 10 mg/kg wasgiven and three male animals for each route were used. All blood sampleswere taken from saphenous vein at 5 min, 30 min, 1 hour, 6 hours, 24hours after intravenous administration and 15 min, 30 min, 1 hour, 6hours, 24 hours after oral administration. Plasma samples were preparedby plasma proteins precipitation adding 200 μL of methanol to 10 μL ofplasma in a 96 well plate. After capping and vortex mixing, the platewas centrifuged for 15 min at 3700 rpm at 6° C. The supernatant wasconsidered as final extract and injected onto the LC-MS-MS system (HPLCsystem: Hewlett Packard 1100 series using Atlantis HILIC Silica 50*2.1mm 5.0 μm analytical column; MS instrument: Perkin Elmer SCIEX API 2000and ionization performed with Turbo Ion Spray in positive ion mode).Lower limit of quantification is 5.0 ng/mL, upper limit ofquantification is 10000 ng/mL. Analysis was performed using the Watsonpackage (version 6.4.0.04) and Excel spreadsheet (Microsoft Inc.Seattle, USA). Non-compartmental method (linear trapezoidal rule andlinear regression analysis of natural log-transformed plasmaconcentrations vs. time data) was used. After intravenous dosing, Co wasset equal to Co 083 Absolute bioavailability (F) was calculated from theratio of average oral to IV (intravenous) dose-normalized plasma AUC(area under curve) values.

Some representative compounds of formula (I) were evaluated for theirpharmacokinetic parameters as reported in table 5 as mean value.

TABLE 5 CL Vdss AUC₀₋₂₄ C-max T½ F on Com- (IV bolus) (IV bolus) (oral)(oral) (oral) AUC pounds mL/min/kg L/Kg μM · hours μM hours % 3 32.63.61 13.5 2.53 3.04 100 4 53.4 6.8 4.93 1.37 2.62 78 13 26.7 4.7 18.33.64 2.9 100 18 47 3.84 7.28 3.9 1.8 92 19 88 3.9 4.28 2.85 0.8 93 21 234.9 7.39 1.2 6.6 56

From the above, it is clear to the person skilled in the art thatcompounds of formula (I) possess good to excellent pharmacokineticsprofiles and oral bioavailability.

In Vivo Efficacy Studies

Balb, athymic Nu/Nu male mice, from Harlan (Italy), were maintained inagreement with the European Communities Council Directive no. 86/609/EECconcerning the protection of animals used for experimental or otherscientific purposes, in cages with paper filter cover, food and beddingsterilized and acidified water. Fragments of Capan-1 human pancreaticcancer tumors were implanted subcutaneously. Mice bearing a palpabletumor (100-200 mm³) were selected and randomized into control andtreated groups. Each group included seven animals. The treatment startedone day after randomization. Compound of formula (I) was administered byoral route as a methocel suspension at the indicated doses and times.Tumor dimension was measured regularly by calipers during theexperiments and tumor mass was calculated as described in Simeoni M. etal., Cancer Res 64, 1094-1101 (2004). The tumor growth inhibition (TGI,%) was calculated according to the equation: % TGI=100-(mean tumorweight of treated group/mean tumor weight of control group)*100.

Some representative compounds of formula (I), were evaluated for theiranti-tumor activity as single agent on Capan-1 BRCA-2 mutated mousemodel. Toxicity was evaluated on the basis of body weight reduction (nobody weight reduction observed out of 7 mice treated). The results arereported in table 6.

TABLE 6 Compounds Dose Schedule Max TGI (%) Toxicity 3 100 mg/kg 1-10daily 51% 0/7 4  75 mg/kg 1-10 daily 52% 0/7 13 100 mg/kg 1-20 daily 18%0/7 21 100 mg/kg 1-20 daily 23% 0/7

Compounds of formula (I) of the present invention, suitable foradministration to a mammal, e.g., to humans, can be administered by theusual routes and the dosage level depends upon the age, weight,conditions of the patient and administration route.

For example, a suitable dosage adopted for oral administration of acompound of formula (I) may range from about 1 to about 1000 mg perdose, from 1 to 5 times daily. The compounds of the invention can beadministered in a variety of dosage forms, e.g., orally, in the formtablets, capsules, sugar or film coated tablets, liquid solutions orsuspensions; rectally in the form suppositories; parenterally, e.g.,intramuscularly, or through intravenous and/or intrathecal and/orintraspinal injection or infusion.

As stated above, the present invention also includes pharmaceuticalcompositions comprising a compound of formula (I) or a pharmaceuticallyacceptable salt thereof in association with a pharmaceuticallyacceptable excipient, which may be a carrier or a diluent.

The pharmaceutical compositions containing the compounds of theinvention are usually prepared following conventional methods and areadministered in a suitable pharmaceutical form. For example, the solidoral forms may contain, together with the active compound, diluents,e.g., lactose, dextrose saccharose, sucrose, cellulose, corn starch orpotato starch; lubricants, e.g., silica, talc, stearic acid, magnesiumor calcium stearate, and/or polyethylene glycols; binding agents, e.g.,starches, arabic gum, gelatine methylcellulose, carboxymethylcelluloseor polyvinyl pyrrolidone; disintegrating agents, e.g., starch, alginicacid, alginates or sodium starch glycolate; effervescing mixtures;dyestuffs; sweeteners; wetting agents such as lecithin, polysorbates,laurylsulphates; and, in general, non-toxic and pharmacologicallyinactive substances used in pharmaceutical formulations. Thesepharmaceutical preparations may be manufactured in known manner, forexample, by means of mixing, granulating, tabletting, sugar-coating, orfilm-coating processes.

The liquid dispersions for oral administration may be, e.g., syrups,emulsions and suspensions. As an example, the syrups may contain, ascarrier, saccharose or saccharose with glycerine and/or mannitol andsorbitol.

The suspensions and the emulsions may contain, as examples of carriers,natural gum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections may contain, together with theactive compound, a pharmaceutically acceptable carrier, e.g., sterilewater, olive oil, ethyl oleate, glycols, e.g., propylene glycol and, ifdesired, a suitable amount of lidocaine hydrochloride.

The solutions for intravenous injections or infusions may contain, as acarrier, sterile water or preferably they may be in the form of sterile,aqueous, isotonic, saline solutions or they may contain propylene glycolas a carrier.

The suppositories may contain, together with the active compound, apharmaceutically acceptable carrier, e.g., cocoa butter, polyethyleneglycol, a polyoxyethylene sorbitan fatty acid ester surfactant orlecithin.

EXPERIMENTAL SECTION

For a reference to any specific compound of formula (I) of theinvention, optionally in the form of a pharmaceutically acceptable salt,see the experimental section and claims. Referring to the examples thatfollow, compounds of the present invention were synthesized using themethods described herein, or other methods, which are well known in theart.

The short forms and abbreviations used herein have the followingmeaning:

μM (micromolar)μL (microliter)μm (micrometer)mol (moles)mM (millimolar)mmol (millimoles)g (grams)mg (milligrams)DC₅₀ (the half maximal Displacement Concentration)IC₅₀ (the half maximal Inhibitory Concentration)

STD (Standard Deviation)

PAR (poly (ADP-ribose))

MEM (Minimal Essential Medium) FCS (Fetal Calf Serum) FBS (Fetal BovineSerum) PBS (Phosphate Buffered Saline) LC-MS (Liquid Chromatography-MassSpectrometry) HPLC (High Performance Liquid Chromatography)

AUC (area under the plasma concentration vs. time curve up to the lastdetectable concentration)Cl (plasma clearance)Cmax (maximum plasma concentration)T½ (terminal half life)Vdss (volume of distribution at steady state)

TGI (Tumor Growth Inhibition)

MHz (megahertz)

Hz (Hertz)

DMSO (dimethylsulfoxide)ESI (electrospray ionization)

With the aim to better illustrate the present invention, without posingany limitation to it, the following examples are now given.

As used herein the symbols and conventions used in the processes,schemes and examples are consistent with those used in the contemporaryscientific literature, for example, the Journal of the American ChemicalSociety or the Journal of Biological Chemistry.

Unless otherwise noted, all materials were obtained from commercialsuppliers, of the best grade and used without further purification.Anhydrous solvent such as N,N-dimethylformamide, tetrahydrofuran,dichloromethane and toluene were obtained from the Aldrich ChemicalCompany. All reactions involving air- or moisture-sensitive compoundswere performed under nitrogen or argon atmosphere.

General Purification and Analytical Methods

Flash Chromatography was performed on silica gel (Merck grade 9395,60A). HPLC was performed on Waters X Terra RP 18 (4.6×50 mm, 3.5 μm)column using a Waters 2790 HPLC system equipped with a 996 Waters PDAdetector and Micromass mod. ZQ single quadrupole mass spectrometer,equipped with an electrospray (ESI) ion source. Mobile phase A wasammonium acetate 5 mM buffer (pH 5.5 with acetic acid-acetonitrile95:5), and mobile phase B was water-acetonitrile (5:95). Gradient from10 to 90% B in 8 minutes, hold 90% B 2 minutes. UV detection at 220 nmand 254 nm. Flow rate 1 mL/min. Injection volume 10 μL. Full scan, massrange from 100 to 800 amu. Capillary voltage was 2.5 KV; sourcetemperature was 120° C.; cone was 10 V. Retention times (HPLC r.t.) aregiven in minutes at 220 nm or at 254 nm. Mass are given as m/z ratio.

When necessary, compounds were purified by preparative HPLC on a WatersSymmetry C18 (19×50 mm, 5 μm) column or on a Waters X Terra RP 18(30×150 mm, 5 μm) column using a Waters preparative HPLC 600 equippedwith a 996 Waters PDA detector and a Micromass mod. ZMD singlequadrupole mass spectrometer, electron spray ionization, positive mode.Mobile phase A was water-0.01% trifluoroacetic acid, and mobile phase Bwas acetonitrile. Gradient from 10 to 90% B in 8 min, hold 90% B 2 min.Flow rate 20 mL/min. In alternative, mobile phase A was water-0.1%ammonium hydroxide, and mobile phase B was acetonitrile. Gradient from10 to 100% B in 8 min, hold 100% B 2 min. Flow rate 20 mL/min.

1H-NMR spectrometry was performed on a Mercury VX 400 operating at400.45 MHz equipped with a 5 mm double resonance probe [1H (15N-31P)ID_PFG Varian].

Example 1 Step a 2-Bromo-4-fluoro-6-methylaniline

A solution of N-bromosuccinimide (18.7 g, 0.105 mol) in 70 mL ofN,N-dimethylformamide was added dropwise to a solution of4-fluoro-2-methylaniline in 70 mL of N,N-dimethylformamide at 20° C. Thereaction mixture was stirred overnight. The dark solution was pouredinto a mixture of water (1000 mL), brine (50 mL) and ethyl acetate (300mL). The mixture was transferred into a separatory funnel, shacked andseparated. The aqueous phase was extracted with ethyl acetate (4×150mL). The combined organic layers were washed with water (5×100 mL),brine (2×100 mL), dried with sodium sulfate, filtered and concentrated.The ¹H NMR spectrum of the crude material showed at least 95% purity.The product was further purified on silica gel pad (eluent system ethylacetate/n-hexane: 1/8). The pure fractions were combined and evaporatedto give 14.9 g of product. The impure fractions were combined,concentrated, redissolved in diethyl ether (30 mL) and extracted with 5%hydrochloric acid (5×10 mL). The acidic phase was basified with aq.potassium hydroxide and extracted with diethyl ether to provide 0.8 g ofthe title compound. Total yield was 15.7 g (77%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.16 (dd, ³J_(H-F)=8.3 Hz, ⁴J_(H-H)=2.9Hz, 1H, Ar), 6.91 (dd, ³J_(H-F)=9.3 Hz, ⁴J_(H-H)=2.9 Hz, 1H, Ar), 4.83(br. s, 2H, NH₂), 2.16 (s, 3H, CH₃).

Step b 2-Bromo-4-fluoro-6-methylbenzonitrile

A solution of potassium cyanide (16.25 g, 0.25 mol) in 20 mL of waterwas added to a suspension of freshly prepared copper(I) chloride (9.5 g,0.096 mol) in 40 mL of water. copper(I) chloride was observed todissolve initially and then a minute amount of precipitate was formed.Toluene (30 mL) was added and the mixture was chilled to 0° C. in thefridge. 2-Bromo-4-fluoro-6-methylaniline (15.7 g, 0.077 mol) was addedto a mixture of 16.5 mL of 36% aqueous hydrochloric acid and 40 mL ofwater. The resulting suspension was heated until a solution formed. Thesolution was chilled to 2° C. with an ice bath and the aminehydrochloride precipitated. A solution of sodium nitrite (5.34 g, 0.078mol) in 15 mL of water was slowly added keeping the reaction mixturetemperature below 5° C. (ice bath). A powder of sodium carbonatedecahydrate was added in small portions to adjust the pH of the reactionmixture to about 7.

A solution of the diazonium salt was slowly added to the cyanocupratereagent keeping the reaction temperature below 5° C. A bright red-orangeprecipitate formed. The reaction mixture was allowed to warm to 20° C.,kept at this temperature for 14 hours. Then it was slowly heated to 70°C. and kept at this temperature for 1 hour. The precipitate dissolvedalmost completely. The reaction mixture was allowed to cool to 20° C.and filtered. Organic phase was separated and the aqueous phase wasextracted with toluene (3×70 mL). The combined organic layers werewashed with water (2×100 mL), brine (2×100 mL), dried with sodiumsulfate, filtered and concentrated. The crude nitrile (13.9 g, 84%)obtained was used without further purification.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.73 (dd, ³J_(H-F)=8.2 Hz, ⁴J_(H-H)=2.1Hz, 1H, Ar), 7.44 (dd, ³J_(H-F)=9.4 Hz, ⁴J_(H-H)=2.0 Hz, 1H, Ar), 2.52(s, 3H, CH₃). ¹³C NMR (300 MHz, d6-DMSO) δ 163.5 (d, ¹J_(C-F)=257 Hz),147.8 (d, ³J_(C-F)=11 Hz), 126.1 (d, ³J_(C-F)=11 Hz), 118.4 (d,²J_(C-F)=27 Hz), 117 (d, ²J_(C-F)=23 Hz), 115.8, 112.7 (d, ⁴J_(C-F)=3Hz).

Step c 2-Bromo-4-fluoro-6-methylbenzamide

2-Bromo-4-fluoro-6-methylbenzonitrile (0.428 g, 2 mmol) was heated in70% aqueous sulfuric acid (2 mL) for 15 hours at 150° C. The reactionmixture was poured into ice and extracted with ethyl acetate (4×2 mL).The organic phase was washed with water (4×2 mL), brine (2×2 mL), driedwith sodium sulfate, filtered and concentrated to give 300 mg of crudeamide. Pure sample was obtained via recrystallization from benzene ¹HNMR (400 MHz, DMSO-d₆) δ ppm 7.89 (br. s, 1H, NH), 7.65 (br. s, 1H, NH),7.41 (dd, ³J_(H-F)=8.6 Hz, ⁴J_(H-H)=2.2 Hz, 1H, Ar), 7.17 (dd,³J_(H-F)=9.8 Hz, ⁴J_(H-H)=2.2 Hz, 1H, Ar), 3.31 (s, 3H, CH₃).

Step d 2-Bromo-4-fluoro-6-methylbenzoic acid

2-Bromo-4-fluoro-6-methylbenzamide (0.9 g, 3.9 mmol) was dissolved in75% aqueous sulfuric acid (4 mL) at 80° C. sodium nitrite (0.5 g, 7.2mmol) was carefully added in small portions during 1 hour. At the end ofaddition a persistent color of nitrogen dioxide appeared. The reactionmixture was chilled to 20° C. and cold water (15 mL) was added to thereaction mixture. The product was extracted with ethyl acetate (6×2 mL).The organic phase was washed with water (4×2 mL), brine (2×2 mL), driedwith sodium sulfate, filtered and concentrated to give 0.879 g (97%) ofpure acid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.7 (br. s, 1H, OH), 7.47 (dd,³J_(H-F)=8.5 Hz, ⁴J_(H-H)=2.4 Hz, 1H, Ar), 7.22 (dd, ³J_(H-F)=9.6 Hz,⁴J_(H-H)=2.2 Hz, 1H, Ar), 2.31 (s, 1H, CH₃). ¹³C NMR (300 MHz,DMSO-d₆+CCl₄) δ ppm 168, 163 (d, ¹J_(C-F)=250 Hz), 138.4 (d, ³J_(C-F)=8Hz), 134 (d, ⁴J_(C-F)=3 Hz), 118.3 (d, ³J_(C-F)=10 Hz), 116.8 (d,²J_(C-F)=24 Hz), 116 (d, ²J_(C-F)=22 Hz), 19.5.

Step e Methyl 2-bromo-4-fluoro-6-methylbenzoate

A mixture of 2-bromo-4-fluoro-6-methylbenzoic acid (1.94 g, 8.33 mmol),anhydrous potassium carbonate (1.72 g, 12.5 mmol), iodomethane (2.36 g,17 mmol) in N,N-dimethylformamide (15 mL) was vigorously stirred for 23hours at 20° C. The suspension was poured into 70 mL of water. A heavyliquid separated. The product was extracted with ethyl acetate (4×25mL). The organic phase was washed with water (5×20 mL), brine (2×20 mL),dried with sodium sulfate, filtered and concentrated to give 2.07 g(100%) of pure ester. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.18 (dd,³J_(H-F)=8.1 Hz, ⁴J_(H-H)=2.4 Hz, 1H, Ar), 6.91 (dd, ³J_(H-F)=9.0 Hz,⁴J_(H-H)=2.2 Hz, 1H, Ar), 3.96 (s, 3H, OCH₃), 2.35 (s, 3H, CH₃).

Operating in an analogous way, the following compound was obtained:

Methyl 2-iodo-4-fluoro-6-methylbenzoate

¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.27 (s, 3H) 3.86 (s, 3H) 7.21-7.27 (m,1H) 7.63 (m, 1H).

Step f7-Bromo-5-fluoro-2-[3-(morpholin-4-yl)propyl]-2,3-dihydro-1H-isoindol-1-one

A mixture of methyl 2-bromo-4-fluoro-6-methylbenzoate (2.07 g, 8.1mmol), N-bromosuccinimide (1.59 g, 8.91 mmol), benzoyl peroxide (98 mg,0.41 mmol) in carbon tetrachloride was heated at reflux for 4 hours. 68mg of benzoyl peroxide were added and the mixture was refluxed foranother 3 hours. The chilled reaction mixture was filtered andevaporated to give 2.9 g of oily residue. The residue was dissolved in20 mL of toluene. N-(3-aminopropyl)-morpholine (1.16 g, 8.1 mmol) andanhydrous potassium carbonate (3.35 g, 24.3 mmol) were added to thesolution and the reaction mixture was heated at 95° C. for 15 hours,chilled, filtered through Celite and concentrated. The product wasrecrystallized from 5 mL of toluene providing 1 g of target product(35%).

¹H NMR (400 MHz, CDCl₃) δ ppm 7.36 (dd, ³J_(H-F)=8.8 Hz, ⁴J_(H-H)=2.2Hz, 1H, Ar), 7.09-7.13 (m, 1H, Ar), 4.35 (s, 1H, CH₂), 3.62-3.74 (m,6H), 2.34-2.49 (m, 6H), 1.87 (quintet, ³J_(H-H)=7.1 Hz, 1H, CH₂ CH₂CH₂).

Operating in an analogous way, the following compound was obtained:

7-Bromo-2-(1-cyclohexylpiperidin-4-yl)-5-fluoro-2,3-dihydro-1H-isoindol-1-one

¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.20 (m, 4H) 1.70 m (10H), 2.29 (br. s.,3H) 2.91 (d, J=10.99 Hz, 2H) 3.82-4.03 (m, 1H) 4.42 (s, 2H) 7.51 (dd,J=8.18, 2.07 Hz, 1H) 7.62 (dd, J=9.03, 2.20 Hz, 1H).

Tert-butyl4-(5-fluoro-7-iodo-1-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.81 (dd, J=2.20, 8.91 Hz, 1H), 7.51(dd, J=2.20, 8.30 Hz, 1H), 4.37 (s, 2H), 4.16 (tt, J=4.15, 11.78 Hz,1H), 4.06 (d, J=14.04 Hz, 2H), 2.84 (br. s., 2H), 1.68-1.78 (m, 2H),1.62 (qd, J=3.80, 12.40 Hz, 2H), 1.42 (s, 9H)

Step p7-Bromo-2-(1-cyclohexylpiperidin-4-yl)-5-fluoro-3-methyl-2,3-dihydro-1H-isoindol-1-one

To a stirred solution of7-bromo-2-(1-cyclohexylpiperidin-4-yl)-5-fluoro-2,3-dihydro-1H-isoindol-1-one(70 mg, 0.177 mmol) and methyliodide (1.1 equiv., 0.195 mmol, 0.012 mL)in anhydrous tetrahydrofuran (3.5 mL) under nitrogen at −78° C., 1Msodium bis(trimethylsilyl)amide in tetrahydrofuran (1.2 equiv., 0.212mL) was slowly added. The reaction mixture was stirred for 90 min andquenched by adding dropwise a 10% ammonium hydroxide solution.Dichloromethane was added, the layers were separated, and the aqueouslayer was extracted twice dichloromethane. Organic layers werecollected, dried and evaporated. Crude (58 mg) was used without furtherpurification.

ESI(+) MS: m/z 409-411 (MH⁺).

Operating in an analogous way, the following compound was obtained:

Tert-butyl4-(6-fluoro-4-iodo-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.78 (dd, J=2.20, 8.79 Hz, 1H),7.54-7.60 (m, J=0.50, 0.50, 0.50, 1.77, 8.36 Hz, 1H), 4.62 (q, J=7.00Hz, 1H), 4.05 (br. s., 2H), 3.85 (tt, J=3.88, 11.99 Hz, 1H), 2.79 (br.s., 2H), 2.07 (qd, J=4.33, 12.55 Hz, 1H), 1.91 (qd, J=4.52, 12.45 Hz,1H), 1.64-1.79 (m, 2H), 1.42 (s, 9H), 1.39-1.47 (m, 3H)

Operating in an analogous way but in excess of alkylating agent (10equiv.) and base (5 equiv.) the following compounds were obtained:

7-Bromo-2-(1-cyclohexylpiperidin-4-yl)-5-fluoro-3,3-dimethyl-2,3-dihydro-1H-isoindol-1-one

ESI(+) MS: m/z 423-425 (MH⁺).

Tert-butyl4-(6-fluoro-4-iodo-1,1-dimethyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.75 (dd, J=2.14, 8.85 Hz, 1H), 7.66(dd, J=2.20, 8.42 Hz, 1H), 4.04 (br. s., 2H), 3.48-3.59 (m, 1H), 2.79(br. s., 2H), 2.34-2.47 (m, 2H), 1.47-1.57 (m, 2H), 1.45 (s, 6H), 1.43(s, 9H)

Step h Tert-butyl4-(4-cyano-6-fluoro-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate

Tert-butyl4-(6-fluoro-4-iodo-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate(240 mg, 0.5 mmol) and copper(I) cyanide (67 mg, 0.75 mmol) weredissolved in N,N-dimethylformamide (2 mL) and the resulting solution wasrefluxed under nitrogen atmosphere for 3 hours. The solvent was removedunder reduced pressure and the residue was diluted with dichloromethaneand washed with 15% ammonium hydroxide. Organic phase was dried oversodium sulfate, filtered and evaporated. Column chromatography(dichloromethane/ethanol:95/5) afforded title compound (174 mg, 93%yield).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.90-7.99 (m, 2H), 4.78 (q, J=6.67 Hz,1H), 4.06 (d, J=11.23 Hz, 2H), 3.86 (tt, J=3.94, 12.05 Hz, 1H), 2.80(br. s., 2H), 2.10 (qd, J=3.97, 12.47 Hz, 1H), 1.95 (qd, J=4.21, 12.35Hz, 1H), 1.73 (t, J=15.07 Hz, 2H), 1.48 (d, J=6.71 Hz, 3H), 1.42 (s, 9H)

Operating in an analogous way, the following compound was obtained:

Tert-butyl4-(4-cyano-6-fluoro-1,1-dimethyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate

¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.03 (dd, J=2.20, 8.42 Hz, 1H), 7.93(dd, J=2.20, 9.15 Hz, 1H), 4.04 (d, J=4.76 Hz, 2H), 3.54-3.68 (m, 1H),2.78 (br. s., 2H), 2.36-2.47 (m, 2H), 1.52-1.59 (m, 2H), 1.50 (s, 6H),1.43 (s, 9H)

Conversion 36-Fluoro-1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carbonitrilehydrochloride

A solution of tert-butyl4-(4-cyano-6-fluoro-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate(2.7 g, 7.5 mmol) in 4M hydrochloric acid in dioxane (18 mL, 75 mmol)was stirred at 50° C. for 2 hours until HPLC analysis revealed thedisappearance of the starting material. The solvent was removed underreduce pressure and the product was dissolved in diethyl ether anddecanted to obtain the title compound (2.09 g, 95%) as its correspondinghydrochloride.

ESI(+) MS: m/z 274 (MH⁺).

Operating in an analogous way, the following compound was obtained:

6-Fluoro-1,1-dimethyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carbonitrilehydrochloride

¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.78 (br. s., 1H), 8.39 (br. s., 1H),8.04 (dd, J=2.20, 8.42 Hz, 1H), 7.95 (dd, J=2.20, 9.15 Hz, 1H),3.66-3.81 (m, 1H), 3.35-3.42 (m, 2H), 2.97 (br. s., 2H), 2.77 (br. s.,2H), 1.75 (br. s., 2H), 1.50 (s, 6H)

Conversion 42-[1-(4,4-Difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carbonitrile

To a suspension of6-fluoro-1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carbonitrilehydrochloride (153 mg, 0.5 mmol) in dichloromethane (5 mL)4,4-difluoro-cyclohexanone (100 mg, 0.75 mmol) sodium acetate (82 mg, 1mmol) and methanol (1 mL) were added. The resulted solution was stirredat room temperature for 5 hours. Then sodium cyanoborohydride (95 mg,1.51 mmol) was added and the mixture was stirred overnight. The solventswere removed under reduce pressure and the residue was dissolved indichloromethane and washed twice with water. The organic phase was driedover anhydrous sodium sulfate and concentrated in vacuo and the residuewas purified by flash chromatography (dichloromethane/methanol 95:5) togive the title compound (90 mg, 46%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.89-7.99 (m, 2H), 4.77 (q, J=6.63 Hz,1H), 3.69 (br. s., 1H), 2.85-2.99 (m, 2H), 2.18-2.31 (m, 2H), 1.46-1.51(m, 3H)

Operating in an analogous way, the following compounds were obtained:

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carbonitrile

ESI(+) MS: m/z 357 (MH⁺).

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carbonitrile

ESI(+) MS: m/z 370 (MH⁺).

Step i2-[1-(4,4-Difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid hydrochloride

2-[1-(4,4-Difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carbonitrile(80 mg, 0.2 mmol) was dissolved in 37% hydrochloric acid (2 mL) and theresulted solution was refluxed for 16 hours. The solvent was removedunder reduced pressure and the residue was diluted with methanol anddecanted in order to obtained the desired product as white solid (80 mg,97%).

ESI(+) MS: m/z 411 (MH⁺).

Operating in an analogous way, the following compounds were obtained:

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid hydrochloride

ESI(+) MS: m/z 375 (MH⁺).

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid hydrochloride

ESI(+) MS: m/z 389 (MH⁺).

2-[1-(4,4-Difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 126

To a solution of2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid hydrochloride (80 mg, 0.2 mmol) in N,N-dimethylformamide (2 mL)hydroxybenzotriazole ammonium salt (61 mg, 0.4 mmol),1-ethyl-3-(3′-dimethylamino) carbodiimide hydrochloric acid salt (77 mg,0.4 mmol) and N,N-diisopropylethylamine (0.14 mL, 1 mmol) were added.The reaction mixture was stirred at room temperature overnight. Thesolvent was removed under reduce pressure and the residue was dissolvedin ethyl acetate. The solution was washed twice with sodium carbonatesaturated aqueous solution, and the organic phase was dried overanhydrous sodium sulfate and concentrated in vacuo. The crude waspurified by flash chromatography (dichloromethane/methanol 96:4) toafford the title compound (60 mg, 73%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.73 (br. s., 1H), 7.87 (dd, J=2.62,10.80 Hz, 1H), 7.84 (br. s., 1H), 7.70-7.76 (m, 1H), 4.79 (q, J=6.75 Hz,1H), 3.71-3.81 (m, 1H), 2.87-3.01 (m, 2H), 1.51 (d, J=6.59 Hz, 3H)

Operating in an analogous way, the following compounds were obtained:

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 127

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.69 (br. s., 1H), 7.87 (dd, J=2.50,10.68 Hz, 1H), 7.84 (br. s., 1H), 7.73 (dd, J=2.20, 7.69 Hz, 1H),4.73-4.83 (m, 1H), 1.51 (d, J=6.59 Hz, 3H).

2-(1-Cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 128

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 7.86 (dd, J=2.50,10.80 Hz, 1H), 7.72 (dd, J=2.50, 7.87 Hz, 1H), 7.35-7.62 (m, 1H),3.25-3.48 (m, 1H), 2.86-2.99 (m, J=3.78 Hz, 2H), 2.53-2.66 (m, J=3.30,12.69 Hz, 2H), 2.17-2.43 (m, 3H), 1.71-1.89 (m, J=8.18, 8.18 Hz, 4H),1.53-1.65 (m, J=10.01 Hz, 2H), 1.51 (s, 6H), 1.18-1.32 (m, 4H),1.02-1.17 (m, 1H).

Example 2 Step n 4-(1-Furan-2-yl-ethylamino)-piperidine-1-carboxylicacid tert-butyl ester

To an equimolar solution of 1-furan-2-yl-ethanone (250 mg, 2.27 mmol)and 4-amino-piperidine-1-carboxylic acid tert-butyl ester (454 mg, 2.27mmol) in dichloromethane (14 mL) 1M titanium(IV) chloride indichloromethane (1.13 mL, 1.13 mmol) and triethylamine (0.31 mL, 2.27mmol) were added. The reaction mixture was stirred under nitrogenatmosphere for 2 days. Then sodium cyanoborohydride (430 mg, 6.8 mmol)in methanol (7 mL) was added dropwise with stirring and the solution wasallowed to stir overnight at room temperature. 35% sodium hydroxide wasadded and the product was extracted with ethyl acetate. The solvent wasremoved under reduce pressure and the crude was purified by flashchromatography (dichloromethane/methanol:95/5) to give the finalcompound as a red oil (406 mg, 61%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.51 (dd, J=0.85, 1.83 Hz, 1H), 6.35(dd, J=1.83, 3.17 Hz, 1H), 6.19 (dt, J=0.66, 3.20 Hz, 1H), 3.91 (q,J=6.88 Hz, 1H), 3.76 (tt, J=4.10, 13.50 Hz, 2H), 2.73 (br. s., 2H),1.65-1.96 (m, 2H), 1.47-1.64 (m, 1H), 1.38 (s, 9H), 1.26 (d, J=6.71 Hz,3H), 0.98-1.15 (m, 2H). ESI(+) MS: m/z 295 (MH⁺).

Operating in an analogous way, the following compounds were obtained:

N-[1-(Furan-2-yl)ethyl]-4-(morpholin-4-yl)aniline

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.50 (dd, J=0.85, 1.83 Hz, 1H), 6.71 (d,J=8.91 Hz, 2H), 6.54 (d, J=8.91 Hz, 2H), 6.32 (dd, J=1.83, 3.17 Hz, 1H),6.15 (dt, J=0.69, 3.14 Hz, 1H), 5.39 (d, J=8.30 Hz, 1H), 4.50 (qd,J=6.71, 7.20 Hz, 1H), 3.64-3.73 (m, 4H), 2.83-2.92 (m, 4H), 1.41 (d,J=6.84 Hz, 3H).

N-[1-(Furan-2-yl)ethyl]-4-(piperidin-1-yl)aniline

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.50 (dd, J=0.79, 1.77 Hz, 1H), 6.69 (d,J=8.67 Hz, 2H), 6.52 (d, J=8.91 Hz, 2H), 6.32 (dd, J=1.83, 3.17 Hz, 1H),6.15 (d, J=3.17 Hz, 1H), 5.35 (d, J=8.42 Hz, 1H), 4.49 (qd, J=6.71, 7.12Hz, 1H), 2.85 (t, J=5.00 Hz, 4H), 1.59 (quin, J=5.40 Hz, 4H), 1.42-1.50(m, 2H), 1.40 (d, J=6.71 Hz, 3H).

N-[1-(Furan-2-yl)ethyl]-4-(4-methylpiperazin-1-yl)aniline

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.50 (dd, J=0.85, 1.83 Hz, 1H), 6.69 (d,J=8.91 Hz, 2H), 6.53 (d, J=8.91 Hz, 2H), 6.32 (dd, J=1.83, 3.17 Hz, 1H),6.14 (dt, J=0.70, 3.23 Hz, 1H), 5.36 (d, J=8.30 Hz, 1H), 4.49 (quin,J=7.11 Hz, 1H), 2.79-2.99 (m, 4H), 2.34-2.47 (m, 4H), 2.19 (s, 3H), 1.40(d, J=6.71 Hz, 3H).

N-[1-(Furan-2-yl)ethyl]-4-(piperidin-1-ylmethyl)aniline

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.52 (dd, J=0.85, 1.83 Hz, 1H), 6.93 (d,J=7.69 Hz, 2H), 6.54 (d, J=8.30 Hz, 2H), 6.33 (dd, J=1.83, 3.17 Hz, 1H),6.18 (dt, J=0.72, 3.20 Hz, 1H), 5.78 (br. s., 1H), 4.56 (quin, J=7.02Hz, 1H), 3.21 (br. s., 2H), 2.25 (br. s., 4H), 1.45 (br. s., 4H), 1.43(d, J=6.71 Hz, 3H), 1.28-1.41 (m, 2H).

(1S)-1-(Furan-2-yl)-N-[(1R)-1-phenylethyl]ethanamine

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.50 (br. s., 1H), 7.24-7.36 (m, 4H),7.15-7.24 (m, 1H), 6.33 (dd, J=1.89, 2.87 Hz, 1H), 6.17 (d, J=2.56 Hz,1H).

(1R)-1-(Furan-2-yl)-N-[(1R)-1-phenylethyl]ethanamine

¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.53 (br. s., 1H), 7.24-7.35 (m, 1H),7.16-7.24 (m, 1H), 6.36 (dd, J=1.65, 2.50 Hz, 1H), 6.09 (d, J=2.56 Hz,1H), 3.56 (q, J=6.59 Hz, 1H), 3.41 (q, J=7.12 Hz, 1H), 1.22 (d, J=6.84Hz, 3H), 1.17 (d, J=6.47 Hz, 3H).

Step o3-(1-Tert-butoxycarbonyl-piperidin-4-yl)-2-methyl-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-ene-6-carboxylicacid

To a solution of 4-(1-furan-2-yl-ethylamino)-piperidine-1-carboxylicacid tert-butyl ester (6.4 g, 21 mmol) in toluene (300 mL) maleicanhydride (2.1 g, 21 mmol) was added. The reaction mixture was refluxedfor 6 hours and then stirred overnight at room temperature. Theprecipitate solid obtained was filtered, washed with diethyl ether anddried to give the desired compound (6.8 g, 82%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.00 (br. s., 1H), 6.58 (d, J=5.86 Hz,1H), 6.45 (dd, J=1.77, 5.80 Hz, 1H), 4.93 (d, J=1.59 Hz, 1H), 4.00 (d,J=12.45 Hz, 2H), 3.89 (q, J=6.55 Hz, 1H), 3.69 (tt, J=3.80, 12.01 Hz,1H), 2.91 (d, J=9.28 Hz, 1H), 2.71 (br. s., 2H), 2.43 (d, J=9.28 Hz,1H), 1.40 (s, 9H), 1.37 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 393 (MH⁺).

Operating in an analogous way, the following compounds were obtained:

3-Methyl-2-[4-(morpholin-4-yl)phenyl]-1-oxo-1,2,3,6,7,7a-hexahydro-3a,6-epoxyisoindole-7-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.14 (br. s., 2H), 7.39 (d, J=9.03 Hz,2H), 7.11 (d, J=9.03 Hz, 2H), 6.96 (d, J=9.03 Hz, 2H), 6.94 (d, J=9.15Hz, 2H), 6.66 (d, J=5.74 Hz, 1H), 6.60 (d, J=5.74 Hz, 1H), 6.52 (dd,J=1.71, 5.86 Hz, 1H), 6.49 (dd, J=1.71, 5.74 Hz, 1H), 5.04 (d, J=1.71Hz, 1H), 5.02 (d, J=1.71 Hz, 1H), 4.82 (q, J=6.59 Hz, 1H), 4.50 (q,J=6.59 Hz, 1H), 3.68-3.79 (m, 8H), 3.14 (d, J=9.15 Hz, 1H), 3.10 (td,J=4.46, 5.77 Hz, 8H), 2.93 (d, J=9.15 Hz, 1H), 2.57 (d, J=9.15 Hz, 1H),2.53 (d, J=9.15 Hz, 1H), 1.36 (d, J=6.71 Hz, 3H), 1.06 (d, J=6.47 Hz,3H).

3-Methyl-1-oxo-2-[4-(piperidin-1-yl)phenyl]-1,2,3,6,7,7a-hexahydro-3a,6-epoxyisoindole-7-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.13 (br. s., 2H), 7.36 (d, J=8.54 Hz,2H), 7.08 (d, J=8.79 Hz, 2H), 6.95 (d, J=7.93 Hz, 4H), 6.66 (d, J=5.74Hz, 1H), 6.60 (d, J=5.74 Hz, 1H), 6.52 (dd, J=1.71, 5.74 Hz, 2H), 6.49(dd, J=1.71, 5.61 Hz, 1H), 6.19 (s, 1H), 5.04 (d, J=1.71 Hz, 1H), 5.02(d, J=1.71 Hz, 1H), 4.80 (q, J=6.55 Hz, 1H), 4.48 (q, J=6.47 Hz, 2H),1.58-1.70 (m, 8H), 1.48-1.57 (m, 4H), 1.36 (d, J=6.71 Hz, 3H), 1.05 (d,J=6.47 Hz, 3H).

3-Methyl-2-[4-(4-methylpiperazin-1-yl)phenyl]-1-oxo-1,2,3,6,7,7a-hexahydro-3a,6-epoxyisoindole-7-carboxylicacid

ESI(+) MS: m/z 384 (MH⁺).

3-Methyl-1-oxo-2-[4-(piperidin-1-ylmethyl)phenyl]-1,2,3,6,7,7a-hexahydro-3a,6-epoxyisoindole-7-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.70 (d, J=5.86 Hz, 1H), 6.62 (d, J=5.74Hz, 1H), 6.55 (dd, J=1.71, 5.86 Hz, 1H), 6.52 (dd, J=1.71, 5.86 Hz, 1H),5.07 (d, J=1.46 Hz, 1H), 5.04 (d, J=1.71 Hz, 1H), 4.99 (q, J=5.94 Hz,1H), 4.74 (q, J=6.63 Hz, 1H), 1.43 (d, J=6.59 Hz, 3H), 1.13 (d, J=6.47Hz, 3H).

Step p1-Methyl-3-oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylicacid

3-(1-Tert-butoxycarbonyl-piperidin-4-yl)-2-methyl-4-oxo-10-oxa-3-aza-tricyclo[5.2.1.0*1,5*]dec-8-ene-6-carboxylicacid (6.6 g, 16.8 mmol) was dissolved in aqueous 37% hydrochloric acid(80 mL) and the resulted solution was refluxed for 3 hours. The solventwas removed under reduced pressure and the residue was diluted withmethanol and decanted in order to obtained the desired product as whitesolid (3.8 g, 82%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 15.84 (br. s., 1H), 8.65-8.97 (m, 2H),8.55 (br. s., 1H), 8.15 (dd, J=0.55, 7.63 Hz, 1H), 7.95 (d, J=7.57 Hz,1H), 7.85 (t, J=7.69 Hz, 1H), 4.98 (q, J=6.59 Hz, 1H), 4.15 (tt, J=3.74,12.19 Hz, 1H), 3.38 (d, J=12.57 Hz, 1H), 2.90-3.14 (m, 2H), 2.35 (qd,J=4.00, 13.20 Hz, 1H), 2.00 (t, J=10.50 Hz, 2H), 1.59 (d, J=6.71 Hz,3H). ESI(+) MS: m/z 275 (MH⁺).

Operating in an analogous way, the following compounds were obtained:

1-Methyl-2-[4-(morpholin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 15.74 (br. s., 1H), 8.19 (d, J=7.81 Hz,1H), 8.02 (d, J=7.57 Hz, 1H), 7.85-7.94 (m, 1H), 7.47 (d, J=9.03 Hz,2H), 7.09 (d, J=9.15 Hz, 2H), 5.50 (q, J=6.71 Hz, 1H), 3.70-3.82 (m,4H), 3.16-3.21 (m, 4H), 1.38 (d, J=6.71 Hz, 3H).

1-Methyl-3-oxo-2-[4-(piperidin-1-yl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.13 (br. s., 2H), 7.36 (d, J=8.54 Hz,2H), 7.08 (d, J=8.79 Hz, 2H), 6.95 (d, J=7.93 Hz, 4H), 6.66 (d, J=5.74Hz, 1H), 6.60 (d, J=5.74 Hz, 1H), 6.52 (dd, J=1.71, 5.74 Hz, 2H), 6.49(dd, J=1.71, 5.61 Hz, 1H), 6.19 (s, 1H), 5.04 (d, J=1.71 Hz, 1H), 5.02(d, J=1.71 Hz, 1H), 4.80 (q, J=6.55 Hz, 1H), 4.48 (q, J=6.47 Hz, 2H),1.58-1.70 (m, 8H), 1.48-1.57 (m, 4H), 1.36 (d, J=6.71 Hz, 3H), 1.05 (d,J=6.47 Hz, 3H).

1-Methyl-2-[4-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid

¹H NMR (400 MHz, DMSO-d₆) δ ppm 15.66 (br. s., 1H), 10.28 (br. s., 1H),8.19 (d, J=7.57 Hz, 1H), 8.02 (d, J=7.57 Hz, 1H), 7.90 (t, J=7.69 Hz,1H), 7.51 (d, J=8.91 Hz, 2H), 7.15 (d, J=9.03 Hz, 2H), 5.52 (q, J=6.67Hz, 1H), 2.85 (d, J=4.39 Hz, 3H), 1.38 (d, J=6.84 Hz, 3H).

1-Methyl-3-oxo-2-[4-(piperidin-1-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxylicacid

ESI(+) MS: m/z 365 (MH⁺).

Conversion 22-(1-Tert-butoxycarbonyl-piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid

To a solution of1-methyl-3-oxo-2-piperidin-4-yl-2,3-dihydro-1H-isoindole-4-carboxylicacid (4 g, 13.2 mmol) in pyridine (15 mL) potassium carbonate (3.6 g,26.5 mol) and methanol (40 mL) were successively added. Thendi-tert-butyl dicarbonate (3.16 g, 14.5 mmol) was added and the reactionmixture was stirred at room temperature for 4 hours until HPLC analysisrevealed the disappearance of the starting material. The solvent wasremoved under reduced pressure and the residue was dissolved indichloromethane. The solution was washed twice with 5% potassiumhydrogen sulfate and the organic phase was dried over anhydrous sodiumsulfate and concentrated in vacuo. The obtained crude wad diluted withdiethyl ether and decanted to obtain the desired product (3.8 g, 78%) asa white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 15.97 (s, 1H), 8.15 (dd, J=0.61, 7.69Hz, 1H), 7.93 (d, J=7.57 Hz, 1H), 7.83 (t, J=7.57 Hz, 1H), 4.98 (q,J=6.63 Hz, 1H), 4.07 (d, J=12.08 Hz, 2H), 4.01 (tt, J=3.85, 12.33 Hz,1H), 2.83 (br. s., 2H), 2.11 (qd, J=4.39, 12.33 Hz, 1H), 1.97 (qd,J=4.46, 12.43 Hz, 1H), 1.82 (t, J=13.12 Hz, 2H), 1.55 (d, J=6.71 Hz,3H), 1.43 (s, 9H). ESI(+) MS: m/z 375 (MH⁺).

Step i″Tert-butyl-4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylatecpd. 1

To an orange solution of2-(1-tert-butoxycarbonyl-piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid (3.8 g, 10.3 mmol) in N,N-dimethylformamide (60 mL)hydroxybenzotriazole ammonium salt (3.15 g, 20.7 mmol),1-ethyl-3-(3′-dimethylamino) carbodiimide hydrochloric acid salt (3.34g, 20.7 mmol) and N,N-diisopropylethylamine (5.3 mL, 30.9 mmol) wereadded. The reaction mixture was stirred at room temperature overnight.The solvent was removed under reduce pressure and the residue wasdissolved in ethyl acetate. The solution was washed twice with sodiumcarbonate saturated aqueous solution, and the organic phase was driedover anhydrous sodium sulfate and concentrated in vacuo. The crude waspurified by flash chromatography (dichloromethane/methanol: 97:3) toafford the title compound (2.86 g, 74%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.60 (br. s., 1H), 8.17 (dd, J=1.10,7.57 Hz, 1H), 7.78 (ddd, J=0.50, 1.10, 7.69 Hz, 1H), 7.72 (t, J=7.60 Hz,1H), 7.65 (br. s., 1H), 4.79 (d, J=6.71 Hz, 1H), 4.06 (d, J=12.00 Hz,2H), 3.96 (tt, J=3.98, 12.07 Hz, 1H), 2.82 (br. s., 2H), 2.10 (qd,J=4.64, 12.45 Hz, 1H), 1.94 (qd, J=4.58, 12.35 Hz, 1H), 1.77 (t, J=12.76Hz, 2H), 1.49 (d, J=6.71 Hz, 3H), 1.42 (s, 9H). ESI(+) MS: m/z 374(MH⁺).

Operating in an analogous way, the following compounds were obtained:

1-Methyl-2-[4-(morpholin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 35

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.37 (br. s., 1H), 8.20 (dd, J=0.85,7.69 Hz, 1H), 7.87 (d, J=7.50 Hz, 1H), 7.79 (t, J=7.69 Hz, 1H), 7.69(br. s., 1H), 7.43 (d, J=9.03 Hz, 2H), 7.05 (d, J=9.15 Hz, 2H), 5.35 (q,J=6.47 Hz, 1H), 3.70-3.81 (m, 4H), 3.16 (dd, J=4.27, 5.61 Hz, 4H), 1.34(d, J=6.71 Hz, 3H).

1-Methyl-3-oxo-2-[4-(piperidin-1-yl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 36

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.41 (br. s., 1H), 8.20 (dd, J=0.73,7.69 Hz, 1H), 7.86 (d, J=7.70 Hz, 1H), 7.79 (t, J=7.65 Hz, 1H), 7.69(br. s., 1H), 7.38 (d, J=9.03 Hz, 2H), 7.02 (d, J=9.03 Hz, 2H), 5.32 (q,J=6.63 Hz, 1H), 3.15-3.22 (m, 4H), 1.59-1.70 (m, 4H), 1.48-1.60 (m, 2H),1.33 (d, J=6.59 Hz, 3H).

1-Methyl-2-[4-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 37

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.39 (br. s., 1H), 8.20 (dd, J=0.85,7.69 Hz, 1H), 7.87 (d, J=7.60 Hz, 1H), 7.79 (t, J=7.63 Hz, 1H), 7.69(br. s., 1H), 7.41 (d, J=9.03 Hz, 2H), 7.04 (d, J=9.15 Hz, 2H), 5.34 (q,J=6.67 Hz, 1H), 3.19 (t, J=4.90 Hz, 4H), 2.48 (t, J=4.64 Hz, 4H), 2.24(s, 3H), 1.33 (d, J=6.71 Hz, 3H).

1-Methyl-3-oxo-2-[4-(piperidin-1-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 38

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.16 (br. s., 1H), 8.18 (dd, J=0.73,7.69 Hz, 1H), 7.88 (d, J=7.60 Hz, 1H), 7.81 (t, J=7.56 Hz, 1H), 7.71(br. s., 1H), 7.56 (d, J=8.18 Hz, 2H), 7.40 (d, J=8.30 Hz, 2H), 5.45 (q,J=6.43 Hz, 1H), 3.46 (s, 2H), 2.35 (br. s., 4H), 1.51 (quin, J=5.50 Hz,4H), 1.38-1.45 (m, 2H), 1.36 (d, J=6.71 Hz, 3H).

Conversion 31-Methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 2

A solution oftert-butyl-4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate(2.8 g, 7.5 mmol) in 4M hydrochloric acid in dioxane (18 mL, 75 mmol)was stirred at 50° C. for 2 hours until HPLC analysis revealed thedisappearance of the starting material. The solvent was removed underreduce pressure and the product was dissolved in diethyl ether anddecanted to obtain the title compound (2 g 95%) as its hydrochloride.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.53 (br. s., 1H), 8.80 (d, J=9.89 Hz,1H), 8.64 (q, J=10.54 Hz, 1H), 8.18 (dd, J=1.10, 7.57 Hz, 1H), 7.80 (d,J=7.60 Hz, 1H), 7.74 (t, J=7.60 Hz, 1H), 7.69 (br. s., 1H), 4.81 (q,J=6.67 Hz, 1H), 4.11 (tt, J=4.01, 12.16 Hz, 1H), 2.91-3.14 (m, J=23.44Hz, 2H), 1.95 (d, J=14.04 Hz, 2H), 1.54 (d, J=6.71 Hz, 3H). ESI(+) MS:m/z 274 (MH⁺).

Conversion 42-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd. 3

To a suspension of1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide(60 mg, 0.19 mmol) in dichloromethane (2 mL) cyclohexanone (147 mg, 0.28mmol) sodium acetate (32 mg, 0.38 mmol) and methanol (0.3 mL) wereadded. The resulted solution was stirred at room temperature for 5hours. Then sodium cyanoborohydride was added and the mixture wasstirred overnight. The solvents were removed under reduce pressure andthe residue was dissolved in dichloromethane and washed twice withwater. The organic phase was dried over anhydrous sodium sulfate andconcentrated in vacuo and the residue was purified by flashchromatography (dichloromethane/methanol: 95:5) to give the titlecompound.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.70 (br. s., 1H), 8.13-8.22 (m, 1H),7.77 (d, J=7.30 Hz, 1H), 7.71 (t, J=7.57 Hz, 1H), 7.65 (br. s., 1H),4.79 (q, J=6.75 Hz, 1H), 3.77 (tt, J=4.06, 12.24 Hz, 1H), 2.92 (t,J=10.38 Hz, 2H), 2.28 (q, J=12.00 Hz, 3H), 2.12 (qd, J=3.90, 12.20 Hz,1H), 1.97 (qd, J=3.72, 11.94 Hz, 1H), 1.75 (br. s., 6H), 1.58 (d,J=12.08 Hz, 1H), 1.50 (d, J=6.71 Hz, 3H), 1.13-1.30 (m, 4H), 0.99-1.13(m, 1H). ESI(+) MS: m/z 356 (MH⁺). Single enantiomers (99% e.e.) havebeen obtained by preparative chiral-HPLC by using Chiralcel OD 50×500 mm20 μm as column system and n-hexane/ethanol/methanol 80:10:10 as eluant.Configuration of the stereogenic center was assigned by comparison withcpd 4 obtained by synthesis starting from optically pure(1S)-1-(furan-2-yl)ethanamine (see example 4).

(1S)-2-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 4

First eluting peak, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.70 (br. s., 1H),8.18 (dd, J=1.04, 7.63 Hz, 1H), 7.74-7.81 (m, 1H), 7.67-7.75 (m, 1H),7.65 (br. s., 1H), 4.79 (q, J=6.55 Hz, 1H), 3.57-3.94 (m, 1H), 2.92 (t,J=10.62 Hz, 2H), 2.19-2.38 (m, 3H), 2.05-2.19 (m, 1H), 1.90-2.05 (m,1H), 1.75 (br. s., 11H), 1.54-1.62 (m, 2H), 1.49 (d, J=6.59 Hz, 3H).ESI(+) MS: m/z 356 (MH⁺).

(1R)-2-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 5

Second eluting peak, ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.70 (br. s., 1H),8.18 (dd, J=1.10, 7.57 Hz, 1H), 7.75-7.80 (m, 1H), 7.68-7.75 (m, 1H),7.65 (br. s., 1H), 4.79 (q, J=6.59 Hz, 1H), 3.67-3.87 (m, 1H), 2.92 (t,J=10.44 Hz, 2H), 2.19-2.39 (m, 3H), 2.12 (qd, J=3.72, 12.02 Hz, 1H),1.88-2.05 (m, 1H), 1.75 (br. s., 6H), 1.53-1.66 (m, 2H), 1.49 (d, J=6.59Hz, 3H), 1.12-1.29 (m, 4H), 1.09 (t, J=7.02 Hz, 1H). ESI(+) MS: m/z 356(MH⁺).

2-[1-(4,4-Difluorocyclohexyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 6

The title compound was prepared as described in Conversion 4 replacing4,4-difluoro-cyclohexanone for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.69 (br. s., 1H), 8.18 (dd, J=1.04,7.51 Hz, 1H), 7.77 (d, J=7.60 Hz, 1H), 7.72 (t, J=7.60 Hz, 1H), 7.65(br. s., 1H), 4.79 (q, J=6.55 Hz, 1H), 3.78 (tt, J=3.95, 12.04 Hz, 1H),2.95 (t, J=9.70 Hz, 2H), 2.25 (q, J=10.62 Hz, 2H), 1.50 (d, J=6.71 Hz,3H). ESI(+) MS: m/z 392 (MH⁺).

2-[1-(1H-Indo)-5-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 7

The title compound was prepared as described in Conversion 4 replacing1H-indole-5-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.00 (br. s., 1H), 10.69 (br. s., 1H),8.17 (dd, J=1.10, 7.57 Hz, 1H), 7.77 (d, J=7.60 Hz, 1H), 7.71 (t, J=7.60Hz, 1H), 7.65 (br. s., 1H), 7.45 (br. s., 1H), 7.34 (d, J=8.30 Hz, 1H),7.30 (t, J=2.69 Hz, 1H), 7.07 (dd, J=1.04, 8.36 Hz, 1H), 6.39 (t, J=2.01Hz, 1H), 4.79 (q, J=6.55 Hz, 1H), 3.71-3.90 (m, 1H), 3.55 (br. s., 2H),2.83-3.07 (m, 2H), 2.12-2.29 (m, J=12.57 Hz, 1H), 1.93-2.13 (m, 3H),1.65-1.84 (m, 2H), 1.50 (d, J=6.71 Hz, 3H). ESI(+) MS: m/z 403 (MH⁺).

1-Methyl-3-oxo-2-(1-{[1-(propan-2-yl)-1H-indol-3-yl]methyl}piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 8

The title compound was prepared as described in Conversion 4 replacing1-isopropyl-1H-indole-3-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.17 (dd, J=0.98,7.57 Hz, 1H), 7.74-7.78 (m, 1H), 7.68-7.73 (m, 1H), 7.61-7.69 (m, 2H),7.46 (d, J=8.42 Hz, 1H), 7.38 (br. s., 1H), 7.12 (t, J=7.45 Hz, 1H),6.97-7.06 (m, J=7.45 Hz, 1H), 4.64-4.86 (m, 2H), 3.78 (br. s., 1H), 3.66(br. s., 2H), 3.02 (br. s., 2H), 1.49 (d, J=6.59 Hz, 3H), 1.46 (d,J=6.71 Hz, 6H). ESI(+) MS: m/z 445 (MH⁺).

1-Methyl-3-oxo-2-[1-(pyridin-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 9

The title compound was prepared as described in Conversion 4 replacingpyridine-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.50 (d, J=3.91 Hz,1H), 8.18 (dd, J=0.98, 7.57 Hz, 1H), 7.75-7.81 (m, 2H), 7.69-7.74 (m,1H), 7.66 (br. s., 1H), 7.47-7.49 (m, 1H), 7.23-7.29 (m, 1H), 4.80 (q,J=6.55 Hz, 1H), 3.75-3.87 (m, 1H), 3.64 (br. s., 2H), 2.85-3.03 (m, 2H),2.07-2.34 (m, 4H), 1.69-1.82 (m, 2H), 1.51 (d, J=6.71 Hz, 3H). ESI(+)MS: m/z 365 (MH⁺).

2-(1-{[4-(Benzyloxy)-1H-indol-3-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 10

The title compound was prepared as described in Conversion 4 replacing4-benzyloxy-1H-indole-3-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.92 (br. s., 1H), 10.69 (br. s., 1H),8.17 (dd, J=1.04, 7.63 Hz, 1H), 7.74-7.80 (m, J=1.02, 7.63 Hz, 1H), 7.71(t, J=7.63 Hz, 1H), 7.65 (br. s., 1H), 7.58 (d, J=7.20 Hz, 2H), 7.42 (t,J=7.45 Hz, 2H), 7.34 (t, J=7.35 Hz, 1H), 7.09 (br. s., 1H), 6.90-7.01(m, 2H), 6.56 (quin, J=4.27 Hz, 1H), 5.19 (s, 2H), 4.76 (q, J=6.51 Hz,1H), 3.73 (br. s., 3H), 2.83-3.05 (m, 2H), 1.59-1.79 (m, 2H), 1.47 (d,J=6.59 Hz, 3H). ESI(+) MS: m/z 509 (MH⁺).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.10 (d, 1H) 1.13-1.29 (m, 4H) 1.50 (d,J=6.59 Hz, 3H) 1.53-1.65 (m, 1H) 1.65-1.83 (m, 4H) 1.91-2.03 (m, 1H)2.07-2.19 (m, 1H) 2.22-2.33 (m, 2H) 2.85-2.98 (m, 2H) 3.71-3.82 (m, 1H)4.75-4.83 (m, 1H) 7.65 (br. s., 1H) 7.69-7.74 (m, 1H) 7.75-7.79 (m, 1H)8.18 (dd, J=7.63, 1.04 Hz, 1H) 10.70 (br. s., 1H)

Probe Experimental Preparation Step u2-Methylamino-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide

To a stirred suspension of2-chloro-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide (XXIV,Lg=Cl) (150 mg, 0.52 mmol) in dimethylformamide (1.4 mL), a 33% solutionof methylamine in ethanol was added (9 mL, 72 mmol). The reactionmixture was stirred at room temperature for 3 hours. The ethanol wasthen evaporated under reduced pressure and the mixture was diluted withdiethyl ether and filtered. A light-yellow solid was then collected,washed with diethyl ether then with cold water and dried.2-Methylamino-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide wasobtained in moderate yield (100 mg, 68%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.68 (s, 1H), 10.33 (br. s., 1H), 8.61(d, J=2.1 Hz, 1H), 8.34 (dd, J=7.9, 1.1 Hz, 1H), 8.26 (d, J=8.2 Hz, 1H),7.89 (ddd, J=8.2, 7.1, 1.4 Hz, 1H), 7.70 (dd, J=8.7, 2.2 Hz, 1H), 7.67(ddd, J=7.9, 7.2, 0.7 Hz, 1H), 7.35 (d, J=8.8 Hz, 1H), 3.68 (s, 2H),2.54 (s, 3H).

Step v(3-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-propyl)-carbamicacid tert-butyl ester

To a stirred suspension of2-chloro-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide (XXIV,Lg=Cl) (50 mg, 0.17 mmol) in N,N-dimethylformamide (2 mL),(3-amino-propyl)-carbamic acid tert-butyl ester (0.09 mL, 0.51 mmol) andtriethylamine (0.036 mL, 0.26 mmol) were added. The reaction mixture wasstirred at room temperature overnight, evaporated to dryness and thenpurified through by preparative HPLC on a Waters X Terra RP 18 (19×250mm, 5 μm) column. Mobile phase A was 0.05% ammoniumhydroxide/acetonitrile:95/5 and mobile phase B wasacetonitrile/water:95/5. Gradient from 10 to 75% B in 15 min. Fractionscontaining the desired compound were dried, affording 32 mg (44% yield)of(3-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-propyl)-carbamicacid tert-butyl ester.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.65 (s, 1H), 10.09 (br. s., 1H), 8.63(d, J=2.0 Hz, 1H), 8.33 (dd, J=7.9, 1.2 Hz, 1H), 8.28 (d, J=8.2 Hz, 1H),8.12-8.12 (m, 1H), 7.88 (ddd, J=8.2, 7.1, 1.4 Hz, 1H), 7.72 (dd, J=8.8,2.0 Hz, 1H), 7.66 (ddd, J=8.0, 7.1, 0.7 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H),6.83 (t, J=5.7 Hz, 1H), 3.47 (br. s., 2H), 3.02 (q, J=6.8 Hz, 2H), 2.68(t, J=7.3 Hz, 2H), 1.63 (quin, J=6.9 Hz, 2H), 1.37 (s, 9H).

(6-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-hexyl)-carbamicacid tert-butyl ester

To a stirred suspension of2-chloro-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide (XXIV,Lg=Cl) (65 mg, 0.23 mmol) in N,N-dimethylformamide (2 mL),(6-amino-hexyl)-carbamic acid tert-butyl ester hydrochloride (175 mg,0.69 mmol) and triethylamine (0.097 mL, 0.69 mmol) were added. Thereaction mixture was stirred at room temperature overnight, diluted withdichloromethane, and the resulting solution was washed with water thenbrine, dried over sodium sulfate and evaporated to dryness. The crudewas purified by preparative HPLC on Waters X Terra RP 18 (19×250 mm, 5μm) column. Mobile phase A was 0.05% ammoniumhydroxide/acetonitrile:95/5 and mobile phase B wasacetonitrile/water:95/5. Gradient from 10 to 75% B in 15 min. Fractionscontaining the desired compound

1-Methyl-2-{1-[(4-methyl-1,3-thiazol-5-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 11

The title compound was prepared as described in Conversion 4 replacing4-methyl-thiazole-5-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.66 (br. s., 1H), 8.90 (s, 1H), 8.17(dd, J=1.04, 7.63 Hz, 1H), 7.77 (d, J=7.63 Hz, 1H), 7.72 (t, J=7.60 Hz,1H), 7.65 (br. s., 1H), 4.79 (q, J=6.47 Hz, 1H), 3.73-3.89 (m, 1H), 3.68(s, 2H), 2.95 (t, J=10.01 Hz, 2H), 2.35 (s, 3H), 1.97-2.29 (m, 4H), 1.76(t, J=11.80 Hz, 2H), 1.50 (d, J=6.59 Hz, 3H). ESI (+)ESI(+) MS: m/z 385(MH⁺).

2-[1-(2,4-Difluorobenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 12

The title compound was prepared as described in Conversion 4 replacing2,4-difluoro-benzaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.18 (dd, J=1.04,7.63 Hz, 1H), 7.78 (d, J=7.60 Hz, 1H), 7.72 (t, J=7.60 Hz, 1H), 7.65(br. s., 1H), 7.10-7.34 (m, 3H), 4.79 (q, J=6.51 Hz, 1H), 3.71-3.89 (m,1H), 3.57 (s, 2H), 2.85-3.02 (m, 2H), 2.00-2.31 (m, 4H), 1.67-1.84 (m,2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 400 (MH⁺).

2-(1-Cyclopentylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 13

The title compound was prepared as described in Conversion 4 replacingcyclopentanone for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.61 (br. s., 1H), 8.18 (dd, J=0.98,7.57 Hz, 1H), 7.79 (d, J=7.60 Hz, 1H), 7.73 (t, J=7.60 Hz, 1H), 7.67(br. s., 1H), 4.80 (q, J=6.43 Hz, 1H), 3.91 (br. s., 1H), 1.51 (d,J=6.59 Hz, 3H). ESI(+) MS: m/z 342 (MH⁺).

1-Methyl-2-{1-[(6-methylpyridin-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 14

The title compound was prepared as described in Conversion 4 replacing6-methyl-pyridine-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.18 (dd, J=1.04,7.63 Hz, 1H), 7.78 (dd, J=1.10, 7.60 Hz, 1H), 7.72 (t, J=7.60 Hz, 1H),7.66 (s, 1H), 7.66 (t, J=7.60 Hz, 1H), 7.27 (d, J=7.69 Hz, 1H), 7.12 (d,J=7.57 Hz, 1H), 4.80 (q, J=6.55 Hz, 1H), 3.82 (br. s., 1H), 3.58 (br.s., 2H), 2.90-3.05 (m, 2H), 2.45 (s, 3H), 1.67-1.85 (m, 2H), 1.51 (d,J=6.59 Hz, 3H). ESI(+) MS: m/z 379 (MH⁺).

2-[1-(Furan-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 15

The title compound was prepared as described in Conversion 4 replacingfuran-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.66 (br. s., 1H), 8.16 (dd, J=1.04,7.51 Hz, 1H), 7.73-7.78 (m, 1H), 7.67-7.73 (m, 1H), 7.61-7.66 (m, 1H),7.58 (s, 1H), 6.36-6.42 (m, 1H), 6.36-6.42 (m, 1H), 6.25-6.31 (m, 1H),6.29 (br. s., 1H), 4.77 (q, J=6.55 Hz, 1H), 3.76 (d, J=14.28 Hz, 1H),3.52 (br. s., 2H), 2.88-2.98 (m, 2H), 2.13-2.24 (m, 2H), 1.98-2.11 (m,5H), 1.68-1.80 (m, 3H), 1.48 (d, 3H). ESI(+) MS: m/z 354 (MH⁺).

1-Methyl-3-oxo-2-{1-[3-(piperidin-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 16

The title compound was prepared as described in Conversion 4 replacing3-piperidin-1-yl-benzaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.18 (dd, J=1.16,7.63 Hz, 1H), 7.75-7.80 (m, 1H), 7.68-7.75 (m, 1H), 7.66 (br. s., 1H),7.15 (t, J=7.81 Hz, 1H), 6.87 (br. s., 1H), 6.81 (d, J=8.67 Hz, 1H),6.73 (d, J=7.45 Hz, 1H), 4.80 (q, J=6.47 Hz, 1H), 3.68-3.89 (m, 1H),3.44 (br. s., 2H), 3.07-3.17 (m, 4H), 2.90-3.00 (m, 2H), 1.97-2.28 (m,4H), 1.69-1.81 (m, 2H), 1.57-1.68 (m, 4H), 1.53-1.57 (m, 1H), 1.51 (d,J=6.71 Hz, 3H). ESI(+) MS: m/z 447 (MH⁺).

1-Methyl-2-(1-{[6-(morpholin-4-yl)pyridin-2-yl]methyl}piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 17

The title compound was prepared as described in Conversion 4 replacing6-morpholin-4-yl-pyridine-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.69 (br. s., 1H), 8.18 (dd, J=1.04,7.63 Hz, 1H), 7.76-7.81 (m, 1H), 7.69-7.75 (m, 1H), 7.66 (br. s., 1H),7.55 (t, J=7.81 Hz, 1H), 6.79 (d, J=7.20 Hz, 1H), 6.68 (d, J=8.30 Hz,1H), 4.74-4.84 (m, 1H), 3.81 (t, J=7.32 Hz, 1H), 3.66-3.72 (m, 4H), 3.48(s, 2H), 3.40-3.45 (m, 4H), 2.90-3.04 (m, 2H), 2.04-2.31 (m, 4H),1.68-1.82 (m, 2H), 1.51 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 450 (MH⁺).

2-(1-Cyclobutylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 18

The title compound was prepared as described in Conversion 4 replacingcyclobutanone for cyclohexanone. ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68(br. s., 1H), 8.18 (dd, J=0.98, 7.57 Hz, 1H), 7.76-7.80 (m, 1H),7.69-7.75 (m, 1H), 7.66 (br. s., 1H), 4.79 (q, J=6.43 Hz, 1H), 3.70-3.88(m, 1H), 2.83-2.96 (m, 2H), 2.66-2.77 (m, 1H), 1.91-2.23 (m, 4H),1.70-1.85 (m, 5H), 1.62 (br. s., 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+)MS: m/z 328 (MH⁺).

1-Methyl-3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 19

The title compound was prepared as described in Conversion 4 replacing1H-pyrrole-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 10.64 (br. s., 1H),8.18 (dd, J=1.10, 7.57 Hz, 1H), 7.75-7.81 (m, 1H), 7.67-7.75 (m, 1H),7.65 (br. s., 1H), 6.64 (br. s., 1H), 5.93 (br. s., 1H), 5.88 (br. s.,1H), 4.78 (q, J=6.59 Hz, 1H), 3.70-3.85 (m, 1H), 3.43 (br. s., 2H),2.86-3.00 (m, 2H), 1.87-2.30 (m, 4H), 1.75 (t, J=8.79 Hz, 2H), 1.49 (d,J=6.59 Hz, 3H). ESI(+) MS: m/z 353 (MH⁺).

1-Methyl-3-oxo-2-[1-(thiophen-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 20

The title compound was prepared as described in Conversion 4 replacingthiophene-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.18 (dd, J=0.98,7.57 Hz, 1H), 7.75-7.80 (m, 1H), 7.68-7.75 (m, 1H), 7.65 (br. s., 1H),7.48 (br. s., 1H), 7.33 (br. s., 1H), 7.08 (br. s., 1H), 4.79 (q, J=6.71Hz, 1H), 3.79 (t, J=11.29 Hz, 1H), 3.52 (br. s., 2H), 2.88-3.00 (m, 2H),1.92-2.27 (m, 4H), 1.67-1.82 (m, 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+)MS: m/z 370 (MH⁺).

2-[1-(Cyclohexylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 21

The title compound was prepared as described in Conversion 4 replacingcyclohexanecarbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.69 (br. s., 1H), 8.18 (dd, J=0.92,7.51 Hz, 1H), 7.75-7.79 (m, 1H), 7.68-7.75 (m, 1H), 7.62-7.68 (m, 1H),4.79 (q, J=6.55 Hz, 1H), 3.65-3.84 (m, 1H), 2.84-2.97 (m, 2H), 1.50 (d,J=6.59 Hz, 3H), 0.74-0.91 (m, 2H). ESI(+) MS: m/z 370 (MH⁺).

2-[1-(Cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd

The title compound was prepared as described in Conversion 4 replacingcyclohex-3-enecarbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.69 (br. s., 1H), 8.18 (dd, J=0.79,7.63 Hz, 1H), 7.76-7.80 (m, 1H), 7.70-7.76 (m, 1H), 7.66 (br. s., 1H),5.60-5.69 (m, 2H), 4.80 (d, J=6.59 Hz, 1H), 3.79 (br. s., 2H), 2.90-3.03(m, 4H), 1.50 (d, J=6.71 Hz, 3H). ESI(+) MS: m/z 368 (MH⁺).

2-{1-[(3-Fluoropyridin-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 23

The title compound was prepared as described in Conversion 4 replacing3-fluoro-pyridine-4-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.52 (d, J=1.59 Hz,1H), 8.41-8.44 (m, 1H), 8.18 (dd, J=1.04, 7.63 Hz, 1H), 7.75-7.80 (m,1H), 7.69-7.74 (m, 1H), 7.66 (br. s., 1H), 7.53 (d, J=11.11 Hz, 1H),4.73-4.83 (m, 1H), 3.80 (d, J=4.52 Hz, 1H), 3.64 (s, 2H), 2.94 (d,J=15.38 Hz, 2H), 2.06-2.35 (m, 5H), 1.69-1.85 (m, 2H), 1.51 (d, J=6.59Hz, 3H). ESI(+) MS: m/z 383 (MH⁺).

1-Methyl-2-{1-[(1-methyl-1H-pyrrol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 24

The title compound was prepared as described in Conversion 4 replacing1-methyl-1H-pyrrole-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.17 (dd, J=1.10,7.57 Hz, 1H), 7.77 (d, J=7.60 Hz, 1H), 7.71 (t, J=7.60 Hz, 1H), 7.65(br. s., 1H), 6.66 (t, J=2.01 Hz, 1H), 5.87 (d, J=1.95 Hz, 2H), 4.78 (d,J=6.59 Hz, 1H), 3.70-3.88 (m, 1H), 3.61 (s, 3H), 3.41 (s, 2H), 2.89-3.02(m, 2H), 2.09-2.26 (m, 1H), 1.92-2.09 (m, 3H), 1.75 (t, J=12.80 Hz, 2H),1.49 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 367 (MH⁺).

1-Methyl-2-{1-[3-(methylsulfanyl)propyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 25

The title compound was prepared as described in Conversion 4 replacing1-(3-methylsulfanyl-propyl)-piperidin-4-one for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.18 (dd, J=1.04,7.63 Hz, 1H), 7.78 (d, J=7.60 Hz, 1H), 7.72 (t, J=7.60 Hz, 1H), 7.65(br. s., 1H), 4.79 (q, J=6.59 Hz, 1H), 3.80 (t, J=11.11 Hz, 1H), 2.96(t, J=10.19 Hz, 2H), 2.38 (t, J=7.32 Hz, 2H), 2.20 (q, J=11.80 Hz, 1H),2.05 (s, 3H), 2.00 (quin, J=12.00 Hz, 3H), 1.75 (t, J=11.00 Hz, 2H),1.70 (quin, J=7.00 Hz, 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 362(MH⁺).

2-[1-(1H-Imidazol-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 26

The title compound was prepared as described in Conversion 4 replacing1H-imidazole-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.84 (br. s., 1H), 10.69 (br. s., 1H),8.18 (dd, J=1.16, 7.63 Hz, 1H), 3.53 (s, 2H), 1.50 (d, J=6.71 Hz, 3H).ESI(+) MS: m/z 354 (MH⁺).

2-{1-[(2-Chloropyridin-3-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 27

The title compound was prepared as described in Conversion 4 replacing2-chloro-pyridine-3-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.32 (dd, J=1.71,4.64 Hz, 1H), 8.18 (dd, J=0.98, 7.57 Hz, 1H), 7.96 (dd, J=1.65, 7.38 Hz,1H), 7.78 (d, J=7.60 Hz, 1H), 7.72 (t, J=7.57 Hz, 1H), 7.66 (br. s.,1H), 7.46 (dd, J=4.82, 7.63 Hz, 1H), 4.80 (q, J=6.63 Hz, 1H), 3.74-3.93(m, J=11.90, 11.90 Hz, 1H), 3.61 (s, 2H), 2.94 (t, J=10.44 Hz, 2H), 1.77(t, J=12.21 Hz, 2H), 1.52 (d, J=6.71 Hz, 3H). ESI(+) MS: m/z 399 (MH⁺).

1-Methyl-3-oxo-2-{1-[3-(1H-pyrazol-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,cpd

The title compound was prepared as described in Conversion 4 replacing3-pyrazol-1-yl-benzaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) ppm 10.68 (br. s., 1H), 8.50 (d, J=2.08 Hz,1H), 8.18 (dd, J=1.04, 7.63 Hz, 1H), 7.81 (s, 1H), 7.76-7.79 (m, 1H),7.75 (d, J=1.59 Hz, 1H), 7.72 (d, J=8.06 Hz, 0H), 7.69-7.74 (m, 1H),7.65 (br. s., 1H), 7.46 (t, J=7.75 Hz, 1H), 7.28 (d, J=7.45 Hz, 1H),6.54 (dd, J=1.83, 2.32 Hz, 1H), 4.80 (q, J=6.51 Hz, 1H), 3.82 (br. s.,1H), 3.59 (s, 2H), 2.90-3.08 (m, 2H), 1.77 (br. s., 2H), 1.51 (d, J=6.71Hz, 3H). ESI(+) MS: m/z 430 (MH⁺).

1-Methyl-2-{1-[(5-methylfuran-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd

The title compound was prepared as described in Conversion 4 replacing5-methyl-furan-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.18 (dd, J=0.85,7.57 Hz, 1H), 7.77 (d, J=7.60 Hz, 1H), 7.71 (t, J=7.60 Hz, 1H), 7.65(br. s., 1H), 6.15 (br. s., 1H), 5.99 (br. s., 1H), 4.78 (q, J=6.71 Hz,1H), 3.78 (br. s., 1H), 3.45 (br. s., 2H), 2.81-3.06 (m, 2H), 2.24 (s,3H), 2.12-2.27 (m, OH), 2.04 (br. s., 3H), 1.75 (br. s., 2H), 1.49 (d,J=6.71 Hz, 3H). ESI(+) MS: m/z 368 (MH⁺).

1-Methyl-3-oxo-2-{1-[(3-phenyl-1H-pyrazol-5-yl)methyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 30

The title compound was prepared as described in Conversion 4 replacing5-phenyl-2H-pyrazole-3-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.17 (dd, J=1.22,7.57 Hz, 1H), 7.77 (d, J=7.60 Hz, 1H), 7.71 (t, J=7.60 Hz, 1H), 7.64(br. s., 1H), 4.79 (q, J=6.88 Hz, 1H), 3.82 (br. s., 1H), 3.42 (s, 2H),2.92-3.11 (m, 2H), 2.11-2.29 (m, 2H), 1.94-2.10 (m, 3H), 1.65-1.86 (m,2H), 1.49 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 430 (MH⁺).

2-{1-[(5-Bromothiophen-2-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 31

The title compound was prepared as described in Conversion 4 replacing5-bromo-thiophene-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.66 (br. s., 1H), 8.18 (dd, J=1.04,7.63 Hz, 1H), 7.75-7.81 (m, 1H), 7.69-7.75 (m, 1H), 7.61-7.69 (m, 1H),7.06 (d, J=3.54 Hz, 1H), 6.84 (d, J=2.93 Hz, 1H), 4.79 (q, J=6.63 Hz,1H), 3.80 (br. s., 1H), 3.69 (br. s., 2H), 2.98 (br. s., 2H), 1.76 (br.s., 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 439 (MH⁺).

2-[1-(3-Hydroxybenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 32

The title compound was prepared as described in Conversion 4 replacing3-hydroxy-benzaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 9.26 (s, 1H), 8.18(dd, J=0.98, 7.57 Hz, 1H), 7.75-7.80 (m, 1H), 7.69-7.75 (m, 1H), 7.65(br. s., 1H), 7.10 (t, J=7.75 Hz, 1H), 6.76 (t, J=1.95 Hz, 1H), 6.72 (d,J=7.69 Hz, 1H), 6.63 (ddd, J=0.73, 2.32, 8.06 Hz, 1H), 4.79 (q, J=6.55Hz, 1H), 3.79 (tt, J=3.81, 11.93 Hz, 1H), 3.41 (br. s., 2H), 2.92 (br.s., 1H), 2.24 (qd, J=4.15, 12.30 Hz, 1H), 2.08 (qd, J=4.15, 12.30 Hz,1H), 2.02 (q, J=12.00 Hz, 2H), 1.75 (t, J=12.39 Hz, 2H), 1.51 (d, J=6.59Hz, 3H). ESI(+) MS: m/z 380 (MH⁺).

2-{1-[2-(Difluoromethoxy)benzyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd

The title compound was prepared as described in Conversion 4 replacing2-difluoromethoxy-benzaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.68 (br. s., 1H), 8.18 (dd, J=0.98,7.57 Hz, 1H), 7.74-7.80 (m, 1H), 7.68-7.74 (m, 1H), 7.65 (br. s., 1H),7.50 (dd, J=1.65, 7.26 Hz, 1H), 7.35 (td, J=1.80, 7.50 Hz, 1H), 7.27(td, J=0.85, 7.45 Hz, 1H), 7.19 (d, J=8.06 Hz, 1H), 7.16 (t, J=75.00 Hz,1H), 4.79 (q, J=6.51 Hz, 1H), 3.82 (br. s., 1H), 3.54 (s, 2H), 2.89-3.03(m, 2H), 1.64-1.87 (m, 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+) MS: m/z 430(MH⁺).

2-(1-{[5-(Hydroxymethyl)furan-2-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 34

The title compound was prepared as described in Conversion 4 replacing5-hydroxymethyl-furan-2-carbaldehyde for cyclohexanone.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.67 (br. s., 1H), 8.17 (dd, J=1.10,7.69 Hz, 1H), 7.75-7.81 (m, 1H), 7.68-7.75 (m, 1H), 7.65 (br. s., 1H),6.21 (br. s., 2H), 5.15 (t, J=5.74 Hz, 1H), 4.78 (q, J=6.63 Hz, 1H),4.36 (d, J=5.49 Hz, 2H), 3.79 (br. s., 1H), 3.49 (br. s., 2H), 2.95 (t,J=9.58 Hz, 2H), 1.66-1.85 (m, 2H), 1.50 (d, J=6.59 Hz, 3H). ESI(+) MS:m/z 384 (MH⁺).

Example 3 Step l+e Methyl 4-fluoro-2-iodo-6-methylbenzoate

4-Fluoro-2-methylbenzoic acid (5.0 gr, 32.4 mmol), palladium(II) acetate(0.364 g, 1.62 mmol), iodobenzene diacetate (12.54 g, 38.92 mmol) andelemental iodine (9.88 g, 38.92 mmol) were dissolved inN,N-dimethylformamide (130 mL) and stirred at 100° C. overnight. Thereaction mixture was cooled to room temperature, diluted withmethyl-tert-butylether and aqueous hydrochloric acid and washed with 10%sodium metabisolfite aqueous solution. Organic phase was treated with 2Naqueous sodium hydroxide. The resulting aqueous phase was treated withaqueous hydrochloric acid up to acidic pH and extracted withmethyl-tert-butylether. The organic phase was washed with brine, driedover sodium sulfate and concentrated. So obtained iodinated benzoic acid(6.9 g, 24.6 mmol) was dissolved in N,N-dimethylformamide (50 mL) andmethyliodide (3 mL, 49.2 mmol) and potassium carbonate (5 g, 36.9 mmol)were added. The reaction mixture was stirred overnight at roomtemperature, diluted with methyl-tert-butylether and washed with water,1M sodium hydroxide, water and brine. Solvent was removed under reducedpressure and the crude was purified by flash chromatography(n-hexane/diethyl ether 100:2) to give title compound (5.9 g, 63%yield).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 2.27 (s, 3H) 3.86 (s, 3H) 7.21-7.27 (m,1H) 7.63 (m, J=8.18, 2.44, 0.40, 0.40, 0.40 Hz, 1H).

Example 4 Step q1-Cyclohexyl-N-[(1S)-1-(furan-2-yl)ethyl]piperidin-4-amine

(1S)-1-(Furan-2-yl)ethanamine (0.55 gr, 5 mmol) and1-cyclohexylpiperidin-4-one (0.91 gr, 5 mmol) were dissolved in drydichloromethane (20 mL) in the presence of triethylamine (2 mL, 15 mmol)and titanium(IV) chloride (1M in dichloromethane, 2.5 mmol, 2.5 mL).Reaction mixture was stirred under nitrogen atmosphere for 16 hours.After dilution with methanol (10 mL), sodium cyanoborohydride was added(0.95 gr, 25 mmol) and the resulting reaction mixture was stirred for 4hours. After dilution with ethyl acetate, the organic phase was washedwith 15% ammonium hydroxide and evaporated to dryness. Title compound(0.72 gr, Y=52%) was isolated by column chromatography(dichloromethane/methanol/7N ammonia in methanol 96:2:2)

¹H NMR (400 MHz, DMSO-d₆) δ ppm 1.26 (s, 3H) 3.84-3.93 (m, 1H) 6.17 (d,J=3.17 Hz, 1H) 6.34 (dd, J=3.17, 1.83 Hz, 1H) 7.51 (dd, J=1.71, 0.85 Hz,1H)

Steps o-p(1S)-2-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid

1-Cyclohexyl-N-[(1S)-1-(furan-2-yl)ethyl]piperidin-4-amine (0.3 gr, 1.08mmol) and maleic anhydride (0.13 gr, 1.3 mmol) in toluene (10 mL) werestirred at reflux for 16 hours. Solvent was evaporated and the residuewas dissolved in aqueous 37% hydrochloric acid (5 mL) and stirred at100° C. for 2 hours. Volatiles were evaporated affording crude titleproduct that was used for the next step without further purification

ESI(+) MS: m/z 357 (MH⁺).

Step i″(1S)-2-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,cpd 4

(1S)-2-(1-Cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxylicacid (0.15 gr, 0.42 mmol), hydroxybenzotriazole ammonium salt (0.13 gr,0.84 mmol) and 1-ethyl-3-(3′-dimethylamino) carbodiimide hydrochloricacid salt (0.16 gr, 0.84 mmol) were dissolved in a mixture oftetrahydrofuran (6 mL) and triethylamine (0.23 mL, 1.68 mmol) andstirred under nitrogen atmosphere for 16 hours. Volatiles wereevaporated and the crude was dissolved in dichloromethane and washedwith 15% ammonium hydroxide. The organic phase was dried over anhydroussodium sulfate and concentrated in vacuo. The crude was purified byflash chromatography (dichloromethane/methanol/7N ammonia in methanol100:3:1) to afford the title compound (0.060 g, 40%) as a white solid.

91% e.e. was evaluated by preparative chiral-HPLC (first eluting peak)in comparison with racemic mixture (Cpd 3) by using Chiralcel OD 50×500mm 20 μm as column system and n-hexane/ethanol/methanol 80:10:10 aseluant. were dried, affording 50 mg (47% yield) of(6-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-hexyl)-carbamicacid tert-butyl ester.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.62 (br. s., 1H), 9.90 (br. s., 1H),8.65 (d, J=1.7 Hz, 1H), 8.33 (dd, J=8.5, 1.2 Hz, 1H), 8.30 (d, J=8.0 Hz,1H), 7.88 (ddd, J=8.3, 7.1, 1.1 Hz, 1H), 7.74 (dd, J=8.7, 1.9 Hz, 1H),7.65 (t, J=7.6 Hz, 1H), 7.31 (d, J=8.7 Hz, 1H), 6.73 (t, J=5.5 Hz, 1H),3.27-3.33 (m, 2H), 2.89 (q, J=6.5 Hz, 2H), 2.55 (t, J=7.0 Hz, 2H),1.97-2.61 (m, 1H), 1.17-1.51 (m, 8H), 1.36 (s, 9H).

Step z2-(3-Amino-propylamino)-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamidehydrochloride

(3-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-propyl)-carbamicacid tert-butyl ester (32 mg, 0.075 mmol) was dissolved indichloromethane (1 mL) and 4N hydrochloric acid in dioxane (1 mL) wasadded. The reaction mixture was stirred at room temperature overnightand then evaporated, affording 32 mg (97% yield) of2-(3-amino-propylamino)-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamidebis-hydrochloride.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.72 (s, 1H), 10.90 (s, 1H), 9.29 (d,J=0.6 Hz, 2H), 8.62 (d, J=2.0 Hz, 1H), 8.35 (dd, J=7.9, 1.2 Hz, 1H),8.22 (d, J=8.2 Hz, 1H), 7.98 (br. s., 3H), 7.91 (ddd, J=8.2, 7.1, 1.3Hz, 1H), 7.69 (dd, J=8.8, 2.2 Hz, 1H), 7.68 (ddd, J=7.9, 7.2, 0.9 Hz,1H), 7.38 (d, J=8.8 Hz, 1H), 4.00 (t, J=4.5 Hz, 2H), 3.11 (br. s., 2H),2.92 (sxt, J=6.4 Hz, 2H), 2.00 (quin, J=7.6 Hz, 2H).

2-(6-Amino-hexylamino)-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamidetrifluoroacetate

(6-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-hexyl)-carbamicacid tert-butyl ester was dissolved in dichloromethane (5 mL) andtrifluoroacetic acid (1.5 mL) was added. The reaction mixture wasstirred at room temperature overnight, evaporated, taken up with diethylether, filtered and the collected solid was dried under vacuum to afford30 mg (65% yield) of2-(6-amino-hexylamino)-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamidebis-trifluoroacetate as a light-yellow solid.

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.72 (s, 1H), 10.67 (s, 1H), 8.93 (dt,J=10.7, 5.4 Hz, 2H), 8.58 (d, J=2.1 Hz, 1H), 8.35 (dd, J=8.1, 1.2 Hz,1H), 8.22 (d, J=8.2 Hz, 1H), 7.91 (ddd, J=8.2, 7.1, 1.4 Hz, 1H), 7.69(ddd, J=7.9, 7.2, 0.7 Hz, 1H), 7.65 (dd, J=9.0, 2.3 Hz, 1H), 7.67 (br.s., 3H), 7.38 (d, J=8.7 Hz, 1H), 3.99 (t, J=5.4 Hz, 2H), 2.95-3.06 (m,2H), 2.72-2.85 (m, 2H), 1.64 (quin, J=7.1 Hz, 2H), 1.53 (quin, J=7.0 Hz,2H), 1.33 (dt, J=6.9, 3.4 Hz, 4H).

Step ii9-Dimethylamino-11,11-dimethyl-1-(3-{methyl-[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-carbamoyl}-propyl)-2,3,4,11-tetrahydro-naphtho[2,3-g]quinoliniumtrifluoroacetate (cmpd. P1)

To a stirred solution of2-methylamino-N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-acetamide (3.65mg, 0.013 mmol) in N,N-dimethylformamide (0.5 mL),N,N-diisopropylethylamine (0.0067 mL, 0.039 mmol) and ATTO 610 NHS ester(XXVII) were added under nitrogen, and the reaction was stirred at roomtemperature for 3 hours. The mixture was then evaporated, and theresulting crude was purified by preparative HPLC on Hypersil (21×250 mm,5 μm) column. Mobile phase A was 0.1% trifluoroaceticacid/acetonitrile:95/5 and mobile phase B was acetonitrile/water:95/5.Gradient from 0 to 70% B in 20 min. Fractions containing the desiredcompound were dried, affording 1.6 mg of9-dimethylamino-11,11-dimethyl-1-(3-{methyl-[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-carbamoyl}-propyl)-2,3,4,11-tetrahydro-naphtho[2,3-g]quinoliniumtrifluoroacetate (cmpd. P1).

MS calculated: 654.3444; MS found: 654.3447

ESI(+) MS: m/z 654 (M⁺).

According to this same methodology, but employing the suitable startingmaterials, the following compounds were prepared:

9-Dimethylamino-11,11-dimethyl-1-[3-(3-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-propylcarbamoyl)-propyl]-2,3,4,11-tetrahydro-naphtho[2,3-g]quinoliniumtrifluoroacetate (cmpd. P2)

MS calculated: 697.3866; MS found: 697.3864

ESI(+) MS: m/z 697 (M⁺).

9-Dimethylamino-11,11-dimethyl-1-[3-(6-{[(6-oxo-5,6-dihydro-phenanthridin-2-ylcarbamoyl)-methyl]-amino}-hexylcarbamoyl)-propyl]-2,3,4,11-tetrahydro-naphtho[2,3-g]quinoliniumtrifluoroacetate (cmpd. P3)

MS calculated: 739.4337; MS found: 739.4333

ESI(+) MS: m/z 739 (M⁺).

1. A compound of formula (I):

wherein R is an optionally substituted linear or branched C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl orheteroaryl group; R₁ and R₂ are independently hydrogen atom, anoptionally substituted linear or branched C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl or heteroaryl group,or, taken together with the ring carbon atom to which they are bonded,form an optionally substituted carbon-carbon double bond, a C₃-C₇cycloalkyl or heterocyclyl group, with the proviso that R₁ and R₂ arenot both hydrogen atoms; R₃ is hydrogen or halogen atom, cyano, nitro oran optionally substituted linear or branched C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl, NHCOR₄,COR₄, NR₅R₆, NR₅COR₄, OR₇, SR₇, SOR₁₀, SO₂R₁₀, NHSOR₁₀, NHSO₂R₁₀,R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl group; R₄ is hydrogen atom or anoptionally substituted linear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl,heterocyclyl, aryl, heteroaryl, NR₅R₇, OR₇, SR₇, R₈R₉N—C₁-C₆ alkyl orR₈O—C₁-C₆ alkyl group; R₅ and R₆ are independently hydrogen atom, or anoptionally substituted linear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl, aryl, heteroaryl,R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆ alkyl group, or R₅ and R₆, taken togetherwith the nitrogen atom to which they are bonded, form an optionallysubstituted heterocyclyl group; R₇ is hydrogen atom, or an optionallysubstituted linear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, heterocyclyl, aryl, heteroaryl, COR₅, SOR₁₀,SO₂R₁₀, R₈R₉N—C₁-C₆ alkyl, R₈O—C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl,aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆ alkyl group, wherein R₅ is asdefined above; R₈ and R₉ are independently hydrogen atom or anoptionally substituted linear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclyl, aryl, heteroaryl, COR₄, orR₈ and R₉, taken together with the nitrogen atom to which they arebonded, form an optionally substituted heterocyclyl group, wherein R₄ isas defined above; R₁₀ is hydrogen atom, or an optionally substitutedlinear or branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, heterocyclyl, aryl, heteroaryl, NR₅R₆, OR₇, R₈R₉N—C₁-C₆ alkyl,R₈O—C₁-C₆ alkyl, wherein R₅, R₆, R₇, R₈ and R₉ are as defined above; ora pharmaceutically acceptable salt thereof.
 2. A compound of formula (I)according to claim 1, characterized in that that R is an optionallysubstituted C₃-C₇ cycloalkyl, heterocyclyl, aryl or heteroaryl group,the optional substituents being one or more halogen, cyano, anoptionally further substituted linear or branched C₁-C₆ alkyl, C₃-C₇cycloalkyl, heterocyclyl, aryl, heteroaryl, C₃-C₇ cycloalkyl-C₁-C₆alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), R₈R₉N—C₁-C₆alkyl or R₈O—C₁-C₆ alkyl, the optional further substituents being one ormore halogen, C₁-C₆ alkyl, aryl or heterocyclyl group, NHCOR₄, NR₅R₆,NR₅COR₄, OR₇, oxo (═O), SR₇, R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl, andR₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are as defined in claim 1, or apharmaceutically acceptable salt thereof.
 3. A compound of formula (I)according to claim 1, characterized in that R is an optionallysubstituted C₃-C₇ cycloalkyl, heterocyclyl, aryl or heteroaryl group,the optional substituents being one or more halogen, cyano, anoptionally further substituted linear or branched C₁-C₆ alkyl, C₃-C₇cycloalkyl, heterocyclyl, aryl, heteroaryl or C₃-C₇ cycloalkyl-C₁-C₆alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), R₈R₉N—C₁-C₆alkyl or R₈O—C₁-C₆ alkyl, the optional further substituents being one ormore halogen, C₁-C₆ alkyl, aryl or heterocyclyl group, NHCOR₄, NR₅R₆,NR₅COR₄, OR₇, oxo (═O), SR₇, R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl;either R₁ is hydrogen atom, and R₂ is an optionally substituted linearor branched C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, or R₁ and R₂, taken togetherwith the ring carbon atom to which they are linked, form an optionallysubstituted carbon-carbon double bond, a C₃-C₇ cycloalkyl orheterocyclyl group, and R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are as defined inclaim 1, or a pharmaceutically acceptable salt thereof.
 4. A compound offormula (I) according to claim 1, characterized in that R is anoptionally substituted C₃-C₇ cycloalkyl, heterocyclyl, aryl orheteroaryl group, the optional substituents being one or more halogen,cyano, an optionally further substituted linear or branched C₁-C₆ alkyl,C₃-C₇ cycloalkyl, heterocyclyl, aryl, heteroaryl, C₃-C₇ cycloalkyl-C₁-C₆alkyl, heterocyclyl-C₁-C₆ alkyl, aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆alkyl group, NHCOR₄, COR₄, NR₅R₆, NR₅COR₄, OR₇, oxo (═O), R₈R₉N—C₁-C₆alkyl or R₈O—C₁-C₆ alkyl, the optional further substituents being one ormore halogen, C₁-C₆ alkyl, aryl or heterocyclyl group, NHCOR₄, NR₅R₆,NR₅COR₄, OR₇, oxo (═O), SR₇, R₈R₉N—C₁-C₆ alkyl or R₈O—C₁-C₆ alkyl group;either R₁ is hydrogen atom, and R₂ is an optionally substituted linearor branched C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl,heterocyclyl, aryl or heteroaryl group, or R₁ and R₂, taken togetherwith the ring carbon atom to which they are linked, form an optionallysubstituted carbon-carbon double bond, a C₃-C₇ cycloalkyl orheterocyclyl group; R₃ is hydrogen, halogen atom, cyano, nitro, anoptionally substituted linear or branched C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, NHCOR₄, NR₅R₆, NR₅COR₄, OR₇, R₈R₉N—C₁-C₆ alkyl orR₈O—C₁-C₆ alkyl, and R₄, R₅, R₆, R₇, R₈, and R₉ are as defined in claim1, or a pharmaceutically acceptable salt thereof.
 5. A compound offormula (I) according to claim 1, characterized in that R is anoptionally substituted heterocyclyl or aryl group, the optionalsubstituents being, one or more, optionally further substituted linearor branched C₁-C₆ alkyl, C₃-C₇ cycloalkyl, heterocyclyl, aryl,heteroaryl, C₃-C₇ cycloalkyl-C₁-C₆ alkyl, heterocyclyl-C₁-C₆ alkyl,aryl-C₁-C₆ alkyl or heteroaryl-C₁-C₆ alkyl group or COR₄, and, theoptional further substituents being one or more halogen, C₁-C₆ alkyl,aryl or heterocyclyl group, OR₇, oxo (═O), SR₇, or R₈O—C₁-C₆ alkylgroup; R₁ and R₇ are independently hydrogen atom or a C₁-C₆ alkyl group,with the proviso that they are not both hydrogen atoms; R₃ is hydrogenor halogen atom; R₄ is OR₇; R₇ is hydrogen atom, optionally substitutedC₁-C₆ alkyl or aryl-C₁-C₆ alkyl group, substituents being one or morehalogen atom; R₈ is hydrogen atom, and R₅, R₆ and R₉ are as defined inclaim 1, or a pharmaceutically acceptable salt thereof.
 6. A compound offormula (I) according to claim 1, characterized in that R R is anoptionally substituted piperidinyl or phenyl group, the optionalsubstituents being, one or more, optionally further substituted methyl,ethyl, propyl, cyclohexyl, cyclopentyl, cyclobutyl, morpholinyl,piperazinyl, pyrazolyl, cyclohexyl-methyl, cyclohexenyl-methyl,piperidinyl-methyl, benzyl, pyridyl-methyl, pyrrolyl-methyl,pyrazolyl-methyl, imidazolyl-methyl, thienyl-methyl, indolyl-methyl,thiazolyl-methyl, furyl-methyl group or COR₄; the optional furthersubstituents being one or more bromine, fluorine, chlorine atom orisopropyl, methyl, phenyl, morpholinyl, or piperidinyl, hydroxy-methylgroup, OR₇, oxo (═O) or SR₇; R₁ and R₂ are independently hydrogen atomor a methyl group, with the proviso that they are not both hydrogenatoms; R₃ is hydrogen or fluorine atom; R₄ is OR₇, R₇ is hydrogen atom,an optionally substituted methyl, tert-butyl or benzyl group, thesubstituents being one or more fluorine atom, and R₅, R₆, R₈, and R₉ areas defined in claim 1, or a pharmaceutically acceptable salt thereof. 7.A compound of formula (I) according to claim 1, selected from the groupconsisting of: tert-butyl4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate,1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,(1S)-2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,(1R)-2-(1-cyclohexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(1H-indol-5-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-(1-{[1-(propan-2-yl)-1H-indol-3-yl]methyl}piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(pyridin-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-{[4-(benzyloxy)-1H-indol-3-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{1-[(4-methyl-1,3-thiazol-5-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2,4-difluorobenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methy)-2-{1-[(6-methylpyridin-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(furan-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-{1-[3-(piperidin-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-(1-{[6-(morpholin-4-yl)pyridin-2-yl]methyl}piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclobutylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(thiophen-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclohexylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(3-fluoropyridin-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{1-[(1-methyl-1H-pyrrol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{1-[3-(methylsulfanyl)propyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(1H-imidazol-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(2-chloropyridin-3-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-{1-[3-(1H-pyrazol-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{1-[(5-methylfuran-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-{1-[(3-phenyl-1H-pyrazol-5-yl)methyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(5-bromothiophen-2-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(3-hydroxybenzyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[2-(difluoromethoxy)benzyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-{[5-(hydroxymethyl)furan-2-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[4-(morpholin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[4-(piperidin-1-yl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[4-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[4-(piperidin-1-ylmethyl)phenyl]-7,3-dihydro-1H-isoindole-4-carboxamide,tert-butyl4-(4-carbamoyl-6-fluoro-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidine-1-carboxylate,6-fluoro-1-methyl-3-oxo-2-(piperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-2-[1-(1H-indol-5-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-3-oxo-2-[1-(pyridin-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-2-{1-[(4-methyl-1,3-thiazol-5-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2,4-difluorobenzyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-2-{1-[(6-methylpyridin-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-2-[1-(furan-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-3-oxo-2-{1-[3-(piperidin-1-yl)benzyl]piperidin-4-yl}-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-2-{1-[6-(morpholin-4-yl)pyridin-2-yl]methyl}piperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclobutylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-3-oxo-2-[1-(1H-pyrrol-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-3-oxo-2-[1-(thiophen-2-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclohexylmethyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-2-{1-[(3-fluoropyridin-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-2-{1-[(1-methyl-1H-pyrrol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-2-[1-(1H-imidazol-2-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(2-chloropyridin-3-yl)methyl]piperidin-4-yl}-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-1-methyl-2-{1-[(5-methylfuran-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(5-bromothiophen-2-yl)methyl]piperidin-4-yl}-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,6-fluoro-2-(1-{[5-(hydroxymethyl)furan-2-yl]methyl}piperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,(1S)-2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,(1R)-2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[4-(piperidin-4-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[4-(piperazin-1-ylmethyl)phenyl]-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{4-[(1-methylpiperidin-4-yl)methyl]phenyl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[3-(1-methylpiperidin-4-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[3-(4-methylpiperazin-1-yl)phenyl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[4-(1-methylpiperidin-4-yl)phenyl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[4-cyclohexyl-3-(4-methylpiperazin-1-yl)phenyl]-1-methyl-3-oxo-2,3-dihydro-1H-2-[1-(cyclohex-3-en-1-ylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(2,2,2-trifluoroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamidehydrate,1-methyl-3-oxo-2-[1-(tetrahydrofuran-3-ylmethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,tert-butyl3-{[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]methyl}azetidine-1-carboxylate,1methyl-2-{1-[2-methyl-2-(piperidin-1-yl)propyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[3-(dimethylamino)-2,2-dimethylpropyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-{1-[(1-ethyl-4,5-dihydro-1H-pyrazol-4-yl)methyl]piperidin-4-yl}-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[1-(2-methylbutyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclopentylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,methyl4-[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]butanoate,1-methyl-3-oxo-2-[1-(2,2,2-trichloroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,ethyl2-{[4-(4-carbamoyl-1-methyl-3-oxo-1,3-dihydro-2H-isoindol-2-yl)piperidin-1-yl]methyl}cyclopropanecarboxylate,2-(1-hexylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-(1-nonylpiperidin-4-yl)-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(3-cyclohexylpropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(3-phenylpropyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(cyclopropylmethyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-heptylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-(1-pentylpiperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(2-phenylethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(3,3-dimethylbutyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-(1-propylpiperidin-4-yl)-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(5-hydroxypentyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-3-oxo-2-[1-(2,2,2-trifluoroethyl)piperidin-4-yl]-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-butylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[1-(2-methylpropyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-{1-[(1-methyl-1H-benzimidazol-2-yl)methyl]piperidin-4-yl}-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2,3-dihydroxypropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2,2-dimethylpropyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2,2-dimethylpent-4-en-1-yl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-ethylpiperidin-4-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,1-methyl-2-[1-(3-methylbutyl)piperidin-4-yl]-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-[1-(2-ethylbutyl)piperidin-4-yl]-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclobutylpiperidin-4-yl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclobutylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2′-(1-cyclohexylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1′-isoindole]-4′-carboxamide,2′-(1-cyclohexylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1′-isoindole]-4′-carboxamide,2′-(1-cyclopentylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1′-isoindole]-4′-carboxamide,2′-(1-cyclopentylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1 isoindole]-4′-carboxamide,2′-(1-cyclobutylpiperidin-4-yl)-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1′-isoindole]-4′-carboxamide,2′-(1-cyclobutylpiperidin-4-yl)-6′-fluoro-3′-oxo-2′,3′-dihydrospiro[cyclopropane-1,1′-isoindole]-4′-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylpiperidin-4-yl)-6-fluoro-1-methylidene-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylazetidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylazetidin-3-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylazetidin-3-yl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclopentylazetidin-3-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,0.2-[1-(4,4-difluorocyclohexyl)piperidin-4-yl]-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1-methyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamideand2-(1-cyclohexylpiperidin-4-yl)-6-fluoro-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindole-4-carboxamide,or a pharmaceutically acceptable salt thereof.
 8. A process for thepreparation of a compounds of the formula (I) as defined in claim 1,which process comprises one of the following sequence of steps: SEQUENCEA): Step a) halogenating a compound of formula (X):

wherein R₁ and R₃ are as defined in claim 1; Step b) cyano-de-aminatingthe resultant compound of formula (IX):

wherein Hal is halogen such as Cl, Br, I and R₁ and R₃ are as definedabove; Step c) hydrolyzing the resultant compound of formula (VIII):

wherein Hal, R₁ and R₃ are as defined above; Step d) hydrolyzing theresultant compound of formula (VII):

wherein Hal, R₁ and R₃ are as defined above; Step e) esterifying theresultant compound of formula (VI):

wherein Hal, R₁ and R₃ are as defined above; Step f) cyclizing theresultant compound of formula (V):

wherein T is C₁-C₆ alkyl or aryl C₁-C₆ alkyl group; R₁ and R₃ are asdefined above with a suitable amine of formula (XIV):H₂N—R  XIV wherein R is as defined in claim 1; Step g) alkylating theresultant compound of formula (IV):

wherein Hal, R, R₁ and R₃ are as defined above; Step h)cyano-de-halogenating the resultant compound of formula (III):

wherein Hal, R, R₁, R₂ and R₃ are as defined above; Step i) hydrolyzinga compound of formula (II):

wherein R, R₁, R₂ and R₃ are as defined above to give a compound offormula (I):

wherein R, R₁, R₂ and R₃ are as defined above; or SEQUENCE B): Step l)halogenating a compound of formula (XI):

wherein R₁ and R₃ are as defined above; Step e and m) esterifying theresultant compound of formula (VI) as defined above and cyanode-halogenating the resultant compound of formula (V) as defined above;Step f′) cyclizing the resultant compound of formula (XII)

wherein T, R₁ and R₃ are as defined above, with a suitable amine offormula (XIV) as defined above; Step g′) alkylating the resultantcompound of formula (XIII):

wherein R, R₁ and R₃ are as defined above; Step i′) hydrolyzing theresultant compound of formula (XIII) as defined above to give a compoundof formula (I) as defined above, or SEQUENCE C): either Step n) areductive amination of a compound of formula (XV):

wherein R₁ and R₃ are as defined above; or Step q) a reductivealkylation of a compound of formula (XX):

wherein R₁ and R₃ are as defined above; Step o) reacting with maleicanhydride under Diels-Alder conditions the resultant compound of formula(XVI):

wherein R, R₁ and R₃ are as defined above; Step p) aromatizing theresultant compound of formula (XVII):

wherein R, R₁ and R₃ are as defined above and either Step i″) amidatingthe resultant compound of formula (XVIII):

wherein R, R₁ and R₃ are as defined above and R₂ is hydrogen atom so asto obtain a compound of formula (I) as defined above wherein R₂ ishydrogen atom, or Step e′) esterifying a compound of formula (XVIII) asdefined above wherein R is as defined above; Step s) N-deprotecting theresultant compound of formula (XXI):

wherein T, R₁ and R₃ are as defined above and R is as defined above;Step t) N-alkylating the resultant compound of formula (XXII):

wherein T, R₁ and R₃ are as defined above and; either Step r)hydrolyzing the resultant compound of formula (XXI) as defined abovewherein R is as defined above; or Step g″) alkylating a compound offormula (XXI) as defined above wherein R is as defined above and Stepr′) hydrolyzing the resultant compound of formula (XXIII):

wherein T, R₁, and R₃ are as defined above and R₂ is not hydrogen atomand then converting the resultant compound of formula (XVIII) wherein R,R₁ and R₃ are as defined above and R₂ is not hydrogen atom operating asdescribed above under step i″) and, if necessary or wanted, converting acompound of formula (I) into a different compound of formula (I) byknown chemical reactions; and/or, if desired, converting a compound offormula (I) into a pharmaceutically acceptable salt thereof orconverting a salt into a free compound of formula (I).
 9. Apharmaceutical composition comprising a therapeutically effective amountof a compound of formula (I), as defined in claim 1, and at least onepharmaceutically acceptable carrier and/or diluent. 10-13. (canceled)14. A pharmaceutical composition according to claim 9 further comprisingone or more chemotherapeutic agents.
 15. A product comprising a compoundof formula (I) as defined in claim 1 or a pharmaceutical compositionthereof comprising a therapeutically effective amount of said compoundand at least one pharmaceutically acceptable carrier and/or diluent, andone or more chemotherapeutic agents, as a combined preparation forsimultaneous, separate or sequential use in anticancer therapy.
 16. Amethod for treating diseases mediated by the PARP-1 protein whichcomprises administering to a mammal in need thereof an effective amountof a compound of formula (I), as defined in claim
 1. 17. The method ofclaim 16 wherein the mammal in need thereof is a human.
 18. (canceled)